alu insertions versus blood group plus protein genetic variability in four amerindian populations

Alu insertions versus blood group plus protein genetic variability in fourAmerindian populations1

J Battilana S L Bonattoy L B Freitas M H Hutz T A Weimer S MCallegari-Jacques M A BatzerDagger K Hill A M Hurtadosect L T Tsuneto

M L Petzl-Erler and F M Salzano

Biosciences Institute Federal University of Rio Grande do Sul Porto Alegre Brazily Biosciences Faculty Pontiregcal Catholic University of Rio Grande do Sul Porto Alegre Brazil

Dagger Department of Biological Sciences Biological Computation and Visualization Center Louisiana StateUniversity Baton Rouge USA

Department of Anthropology University of New Mexico Albuquerque USA Department of Genetics Federal University of ParanaAcirc Curitiba Brazil

Received 22 February 2001 in revised form 5 July 2001 accepted 27 July 2001

Summary Background Do the population relationships obtained using DNA or bloodgroup plus protein markers remain the same or do they reveal diŒerent patterns indicatingthat the factors which inmacruence genetic variation at these two levels of analysis are diverseCan these markers shed light on the biological classiregcation of the AcheAcirc a Paraguayan tribewhich only recently established more permanent contacts with non-IndiansSubjects and methods To consider these questions we typed 193 individuals from fourAmerindian tribes in relation to 12 Alu polymorphisms (regve of them never studied inthese populations) while 22 blood group plus protein systems were studied among theAcheAcirc These data were then integrated with those previously available (blood groups plusproteins) for the three other populations DNA extraction and ampliregcation as well as theother laboratory procedures were performed using standard methods currently in use in ourlaboratory The genetic relationships were obtained using the DA distance and the treeswere constructed by the neighbour-joining method both developed by M Nei and colla-borators Reliability of the trees was tested by bootstrap replications Other populationvariability values were also determined using Neirsquos methodsResults Alu polymorphism was observed in all populations and for most of the loci in theseven systems from which we could compare our results with those of other Amerindiangroups agreement was satisfactory Unusual regndings on the blood group plus proteinsystems of the AcheAcirc were a very low (5) HP1 frequency and the presence of the CW

phenotype in the Rh blood group The intertribal patterns of relationship and other aspectsof their variation were remarkably congruent in the two sets (Alu blood group plus protein)of systemsConclusions The answer to the regrst question posed above is a rmative However theproblem of whether the AcheAcirc derived from a GeAtilde group that preceded the Guarani coloniza-tion of Paraguay or are just a diŒerentiated Guarani group could not be answered with thegenetic information available the second hypothesis seems more likely at present but thepoint to be emphasized is the striking genetic distinctiveness of the AcheAcirc as compared toother Amerindians

1 IntroductionAlu insertions the most abundant and extensively studied mammalian class of

SINEs (short interspersed elements) of repetitive sequences represent approximately10 of nuclear DNA in humans (Smit 1996) They are dimeric sequences ofapproximately 300 base pairs (bp) in length (Houck Rinehart and Schmid 1979)originated from the 7SL RNA gene (Ullu and Tschudi 1984) Alu sequences are

ANNALS OF HUMAN BIOLOGY 2002 VOL 29 NO 3 334plusmn347

Annals of Human Biology ISSN 0301plusmn4460 printISSN 1464plusmn5033 online 2002 Taylor amp Francis Ltdhttpwwwtandfcoukjournals

DOI 101080 0301446011008683 5

1 This paper is dedicated to our colleague Maria Helena L P Franco who collaborated in this and other investigations performed by the

Porto Alegre group and whose untimely death is mourned by all of us

postulated to be retrotransposons that have been inserted into the human genomevia a single-stranded RNA intermediate generated by RNA polymerase III tran-scription (Weiner Deininger and Efstratiadis 1986)

There are approximately 2000 Ya58 and 2000 Yb8 insertional elements randomlydispersed throughout the human genome Many of these remacrect recent retropositionevents that have not yet been regxed within the human species (Batzer and Deiniger1991 Batzer Stoneking Alegria-Hartmann et al 1994 Batzer Rubin Hellmann-Blumberg et al 1995 Novick Novick Yunis et al 1995 Arcot De Angelis Sherryet al 1997 Stoneking Fontius CliŒord et al 1997 Novick Novick Yunis et al1998) Due to changes during the evolution of the source genes there are at least 12major Alu subfamilies which may be classireged as old intermediate or young Thetwo oldest subfamilies Jo and Jb arose from independent retroposition eventsinvolving a single ancestral source gene that occurred early in primate evolution(Kapitonov and Jurka 1996) The polymorphic Alu elements may serve as markers toelucidate various aspects of human evolution There are many advantages for theiruse as genetic markers in human populations It is highly improbable that a speciregcAlu sequence has retroposed into a particular site more than once in human evolu-tion and recent insertions represent stable polymorphisms that are rarely lost with-out leaving a trace Because of these two properties identity by descent andevidential stability due to incomplete deletion Alu insertions are literally molecularfossils Another attractive feature of Alu-based population studies is that the ances-tral condition is deregned by the lack of an Alu element Knowledge of the originalcharacter state is useful in rooting phylogenies A further advantage of Alu geneticmarkers is that Alu genotypes involving individual or multiple polymorphic loci canbe determined by a rapid PCR-based assay

For many decades there have been extensive investigations on the biologicalcharacteristics of South Amerindians (Salzano and Callegari-Jacques 1988)Although in recent years the studies have included investigations at the DNAlevel most of the data have been gathered from morphological characteristics ortraditional genetic markers such as blood groups and proteins Despite the consid-erable eŒorts employed the patterns of genetic variability among South Americannative populations are not yet su ciently clear (ClariaAcirc Demarchi Moreno Azoreroet al 1998)

The present study furnishes new information on 12 Alu insertion polymorphismsfor four Amerindian groups as well as blood group and protein genetic data for oneof them (the AcheAcirc of Paraguay) Previous results concerning these latter systemswere compiled for the three other populations and the following questions wereaddressed (a) Are the patterns of relationships involving these four groups thesame considering the two sets of characteristics and (b) Can they provide informa-tion about how the AcheAcirc should be classireged There are competing hypotheses in thisregard some scholars considering them a GeAtilde group that preceded the Guaranicolonization of Paraguay while others contend that they are just a diŒerentiatedGuarani group

2 Subjects and methodsGeneral information about the four populations studied is presented in table 1

and reggure 1 They live in the centralplusmnsouthern part of the continent in Paraguay(AcheAcirc ) and Brazil (the other three) speak languages classireged in the Tupi and GeAtilde plusmnKaingan linguistic groups and the degree of non-Indian socio-cultural inmacruence

Genetic markers in Amerindians 335

received range from minimal (AcheAcirc ) to moderate (Xavante) and extensive(Caingang Guarani) Non-Indian ancestry however was estimated to be verysmall (0plusmn7) for all of them Selected relatively recent additional informationcan be found in Hill and Hurtado (1996) for the AcheAcirc Petzl-Erler Luz andSotomaior (1993) and Salzano Callegari-Jacques Weimer et al (1997a) forthe Caingang and Guarani and Salzano Franco Weimer et al (1997b) for theXavante

The blood samples were collected with anticoagulant refrigerated shortly after-wards and transported in this condition to Porto Alegre or Curitiba The AcheAcirc

samples were submitted to blood group typing immediately after arriving at thelaboratory using the DiaMed micro typing system of gel centrifugation and follow-ing the manufacturerrsquos instructions Haemoglobins were typed by horizontal starch

336 J Battilana et al

Figure 1 Geographic location of the four groups for which genetic data are reported here ACHAcheAcirc CAI Caingang GUA Guarani XAV Xavante

gel electrophoresis employing the method described in Salzano and Tondo (1968)

erythrocyte enzymes according to the techniques given in Harris and Hopkinson(1976) and serum proteins also using horizontal starch gel electrophoresis employ-

ing the buŒer systems of Poulik (1957) for haptoglobin and transferrin and of

Bowman and Bearn (1965) for the other two Amido black 10B was used to stain

albumin and transferrin benzidine to stain haptoglobin and orthodianisidine for

ceruloplasminDNA was extracted following standard procedures (Miller Dykes and Polesky

1988 Lahiri and Nurnberger 1991) The oligonucleotide primers used for the Alu

studies as well as their annealing temperatures have been described by Arcot

Fontius Deininger et al (1995a) Arcot Wang Weber et al (1995b) Arcot

Adamson Lamerdin et al (1996) and Batzer Arcot Phinney et al (1996)

Polymerase chain reaction (PCR) ampliregcation was carried out following Batzerand Deininger (1991) using a regnal volume of 25 mL Each sample was subjected to

the following ampliregcation procedures 1 min at 94degC (denaturation) 2 min at the

appropriate annealing temperature 2 min at 72degC (extension) plus 5 min at 72degC(regnal extension) for 40 cycles Fifteen microlitres of the PCR products were electro-

phoresed in 2 1 TEB (Tris 90 mM Acid Boric 90 mM EDTA 05 M pH 80

0002 mM H2O) agarose gels containing ethidium bromide and the reaction prod-ucts were directly visualized using ultraviolet macruorescence

Allele frequencies were computed either by gene counting or using the MAXLIK

program described by Reed and Schull (1968) A chi-square test for goodness of regt

was used to verify if the observed genotype frequencies agreed with those expected

under HardyplusmnWeinberg equilibrium using the Arlequin program (Schneider

KueŒer Roessli et al 1997) Inter-population heterogeneity was tested by the chi-square test of RoŒand Bentzen (1989) for small numbers


Table 1 Characterization of the groups studied

Population

Characteristics AcheAcirc (Guayaki) Caingang Guarani Xavante

Localities Arroyo Bandera Nonoai Amambai EteAcirc nAuml iteAcirc paChupa-pou LimaAuml o Verde

Porto LindoRio das Cobras

Geographical 55deg50 0W 23deg30 0S 52deg45 0W 27deg20 0S 55deg12 0W 23deg6 0S 51deg40 0W 13deg20 0Scoordinates 56deg30 0W 24deg10 0S 55deg6 0W 23deg12 0S

54deg30 0W 23deg48 0S52deg30 0W 25deg20 0S

Language Guayaki Kaingang Guarani ChavanteLinguistic group Tupi GeAtilde plusmnKaingan Tupi GeAtilde plusmnKainganNon-Indian minimal extensive extensive moderate

socio-culturalinmacruence

Year of sample 1997 1975 2000 1990 1998 1990collection

Non-Indian 0 7 3 2ancestry ()y

According to Greenberg (1987)y According to Salzano and Callegari-Jacques (1988) and Callegari-Jacques and Salzano (1999) usingSzathmary and Reedrsquos (1978) method

The genetic relationships between the four populations were evaluated using theDA genetic distance of Nei Tajima and Tateno (1983) These authors demonstrated

that this method shows a good performance in all tree-making procedures generallygiving a larger probability value and a smaller deviation from the true values in

simulation experiments It also shows a better performance than others for closelyrelated populations such as those of humans Trees were constructed by theneighbour-joining method (Saitou and Nei 1987) Additionally three-dimensional

plots based on principal coordinate and principal components analysis were alsoobtained (Sneath and Sokal 1973) The reliability of the trees was tested by bootstrapreplications (Hedges 1992) For the genetic distances calculations the DISPAN

(Ota 1993) and NTSYS (Rohlf 1987) programs were used Average heterozygosities associated standard errors and other population variability values were determined

according to Nei (1987)

3 Results and discussionInformation about the presence of the 12 Alu insertions in the four population

groups is presented in table 2 Due to a series of circumstances (DNA availability orquality) sample sizes varied in relation to the sites studied (AcheAcirc 31plusmn75 Caingang40plusmn50 Guarani 24plusmn35 Xavante 29plusmn33) but on the whole they can be considered as

fairly representative of the groups under study Polymorphism was observed in allpopulations and for most of the loci Exceptions are 475 regxed in all tribes APO

uniformly present in the AcheAcirc and Xavante ACE in the Ache and FXIIIB in theXavante A25 432 and 465 showed the lowest insertion frequencies 465 beingcompletely absent in the AcheAcirc Most distributions were in HardyplusmnWeinberg equi-

librium Some of them however showed departures from it This is not unexpectedsince in these small endogamous communities mating is far from random Also due

to the number of comparisons made it is expected that some may show such patternjust by chance and regnally due to the small sample sizes the position of just a few

individuals in a given category may greatly inmacruence the probability valuesExtensive repeated tests were performed in the samples with the unexpected distri-butions conregrming the typings previously obtained

For regve insertions (475 323 459 465 432) these are the regrst results obtainedin Amerindians For the other seven the number of populations and individualssampled is variable and a summary is given in table 3 TPA25 (16 populations

593 individuals) is the most studied in North and Central America while forSouth America APO (respectively 19 and 713) is the most studied Generally our

results in these seven systems agree with those obtained elsewhere the largest dif-ference occurring in A25 (our average 9 North plus Central America average21) This also agrees with the fact that as is shown in table 3 the diŒerences in

prevalences in the north plus central and south areas are also smallTwenty-two blood group and protein systems were investigated among the AcheAcirc

and the results are presented in table 4 They were monomorphic for 13 of them(ABO Kell Lutheran haemoglobin (two loci) glucose-6-phosphat e dehydrogenasephosphogluconate dehydrogenase phosphoglucomutas e 2 adenylate kinase acid

phosphatase transferrin ceruloplasmin albumin) in accordance with previousstudies performed in South American Indians Unusual regndings were the low(5) HP1 frequency and the presence of the CW phenotype in the Rh blood group



Table

2P

rese

nce

of

Alu

inse

rtio

ns

am

ong

mem

ber

sof

four

South

Am

eric

an

Ind

ian

popu

lati

on

s

Ach

eAcircC

aingan

gG

uar

ani

Xav

ante

Alu

No

of

Allel

eN

o

of

Allel

eN

o

of

Allel

eN

o

of

Allel

ein

sert

ion

Gen

oty

pe

indiv

iduals

freq

uen

cies

indiv

iduals

freq

uen

cies

indiv

iduals

freq

uen

cies

indiv

idu

als

freq

uen

cies

FX

IIIB

DaggerDagger

45

0782

320

872

2709

35

3010

00

Dagger21

114

05

00

0H

Weq

p

02

96

0001

p10

00

00

00

p10

0000

00

NA

475

DaggerDagger

74

1000

401

000

2710

00

2910

00

Dagger0

00

00

00

0H

Weq

N

AN

AN

AN

A

MA

BD

1Dagger

Dagger10

0581

320

706

903

94

1005

32

Dagger16

18

135

1316

8H

Weq

p

10

00

0000

p00

00

00

00

p00

0900

00

p

04

7100

01

323

DaggerDagger

61

0908

90

420

2008

00

1707

12

Dagger7

248

133

172

3H

Weq

p

00

08

0000

p10

00

00

00

p03

0800

01

p

10

0000

00

A25

DaggerDagger

00

013

00

037

000

97

102

34

Dagger2

36

1373

3825

18H

Weq

p

10

00

0000

p10

00

00

00

p10

0000

00

p

06

5200

01

TP

A25

DaggerDagger

34

0866

170

675

1707

10

604

17

Dagger3

2010

134

34

11H

Weq

p

00

01

0000

p04

87

00

02

p02

1900

01

p

07

0500

01

340 J Battilana et al45

9Dagger

Dagger29

09

68

31

08

54

16

06

5215

0717

Dagger2

811

13

02

62

HW

eq

p

1000

00

00p

0181

0001

p

0137

0001

p

10

00

0000

46

5Dagger

Dagger0

00

00

201

59

000

210

0078

Dagger0

91

531

30

23

27

HW

eq

NA

p

0240

0001

p

1000

0000

p

10

00

0000

AP

ODagger

Dagger36

10

00

37

09

63

32

09

4133

1000

Dagger0

30

00

02

0H

Weq

N

Ap

1000

0000

p

0001

0000

NA

43

2Dagger

Dagger7

01

98

202

50

101

302

0242

Dagger11

20

511

45

26

21

18

HW

eq

p

0001

00

00p

0700

0001

p

0356

0001

p

10

00

0000

PV

92

DaggerDagger

51

08

55

28

07

93

20

07

8323

0813

Dagger16

97

62

43

3H

Weq

p

0619

00

01p

0449

0001

p

0100

0001

p

00

47

0001

AC

EDagger

Dagger76

10

00

17

05

43

25

08

2915

0683

Dagger0

16

811

013

24

HW

eq

NA

p

0043

0001

p

0235

0001

p

04

17

0002

T

he

pre

sen

cean

dabse

nce

of

the

Alu

repea

tar

eden

ote

dby

Daggerand

re

spec

tivel

y

HW

eq

Exac

tte

stfo

rH

ardyplusmnW

einber

geq

uilib

rium

(pro

babilit

yva

lue

plu

sor

min

us

its

standard

erro

r)

NA

N

ot

applica

ble

Three other blood group and protein studies had been conducted among the Ache

(or Guayaki a term that the Ache consider as derogatory) They have been reported

by Matson Sutton Swanson et al (1968) Brown Gajdusek Leyshon et al (1974)

and ClariaAcirc et al (1998) and involved all southern while our data were obtained innorthern groups Respectively 16 15 and 4 allele distributions could be compared

considering the present data and those obtained by these authors and signiregcantdiŒerences were observed in the MNSs Rh Lewis and haptoglobin systems only

Generally the most marked diversity was found between the frequencies reported

here and those given by Brown et al (1974) We have found high (53 vs 28)

LNs (and complementary low LMs) as well as high (82 vs 60) RHCDe

(compensated by low RHcDE) frequencies as compared to those obtained by theindicated authors In the Lewis system our regnding of 67 only of LEWISLe

against 100 found by Matson et al (1968) is much more in accordance with

other studies using this marker (Salzano and Callegari-Jacques 1988) Finally as

far as we can ascertain the AcheAcirc HP1 frequency observed here is the lowest

reported so far in South American Indians In the other studies with this group

previously mentioned the values were also low (22 reported both by Matson

et al 1968 and Brown et al 1974 against a general average for South AmericanIndians of 60 14 Santos Ribeiro-dos-Santos Guerreiro et al 1998) but not

as low as the present reggure Repeated typings were also performed to conregrm our

regndings and they provided uniformly the same result

Are the Alu diŒerences found among the four tribes under consideration con-

gruent with those observed for the blood group plus protein systems To answer this

question we assembled previous results available for the latter (displayed in the

Appendix) to establish the comparison These data were obtained from the samecommunities studied here with the exception of the Guarani who had been pre-

viously surveyed in a more southerly place (see reggure 1 and table 1)

The two sets of genetic distances are shown in table 5 and the derived dendro-

grams in reggure 2 As can be seen they depict essentially the same picture the

Caingang and Xavante (who speak languages of the same linguistic group) clustering


Table 3 Characteristics of previous studies involving seven Alu insertions performed in Amerindians

LociGeographical regionand statistical characteristics FXIIIB MABD1 A25 TPA25 APO PV92 ACE

North and Central AmericaNo of samples 10 2 2 16 10 10 10No of individuals 323 101 101 593 323 323 323Lowest frequency 050 045 021 029 090 057 044Highest frequency 100 046 021 066 100 099 089Mean 084 045 021 055 097 075 070

South AmericaNo of samples 17 4 ETH 21 19 19 17No of individuals 665 313 ETH 668 713 704 508Lowest frequency 053 042 ETH 012 058 042 057Highest frequency 100 065 ETH 093 100 100 098Mean 090 054 ETH 053 097 087 079

Sources Batzer et al (1994) Barley Blackwood Carter et al (1994) TishkoŒ Ruano Kidd et al (1996)Stoneking et al (1997) Novick et al (1998) Oliveira (1999) Rupert Devine Monsalve et al (1999)TishkoŒ Pakstis Stoneking et al (2000)


Table 4 Blood group and protein genetic systems studied among the AcheAcirc

No of Alleles or HWSystem Phenotype individuals haplotypes Frequency Agrave2

ABO O 99 ABOO 1000 NA

MNSs MS 1 LMS 0222 2509MSs 3 LMs 0247 3 dfMs 7 LNs 0531 p lt 0001MNS 0MNSs 39MNs 32NS 0NSs 0Ns 17

P P1 97 P1 0858 NAP2 2

Rh CCwDe 1 RHCDE 0011 295CDE 1 RHCDe 0813 2 dfCDEe 0 RHCwDe 0005 p gt 020CDe 66 RHcDE 0105CcDE 0 RHcDe 0066CcDEe 16CcDe 12cDE 2cDEe 1cDe 0

Kell K 99 KELLK 1000 NA

DuŒy aDaggerbDagger 34 FyA 0828 425aDaggerb 65 1 df

p lt 005

Lewis a bDagger 88 LEWISLe 0667 NAa b 11

Lutheran a bDagger 99 Lub 1000 NA

Haemoglobin A 99 HBA 1000 NAA2 99 HBA2 1000 NA

Glucose-6-phosphate- B 45 F G6PDB 1000 NAdehydrogenase 54 M

Phosphogluconate A 99 PGDA 1000 NAdehydrogenase

Phosphoglucomutase 1 1-1 48 PGM11 0707 0532-1 44 1 df2-2 7 p gt 030

Phosphoglucomutase 2 1-1 99 PGM21 1000 NA

Phosphoglycolate 1-1 62 PGP1 0788 011phosphatase 2-1 32 1 df

2-2 5 p gt 070

Adenylate kinase 1-1 99 AK1 1000 NA

Acid phosphatase B 99 ACPB 1000 NA

Esterase D 1-1 83 ESD1 0919 0762-1 16 1 df2-2 0 p gt 030

Glyoxalase 1 1-1 5 GLO1 0364 12352-1 62 1 df2-2 32 p lt 0001

Haptoglobin 1-1 1 HP1 0050 2452-1 8 1 df2-2 90 p gt 010

Transferrin C 99 TFC 1000 NACeruloplasmin B 99 CPB 1000 NAAlbumin A 99 ALBA 1000 NA

HW HardyplusmnWeinberg (chi-square test for equilibrium) NA Not applicable

together In both cases the AcheAcirc remain far from the others with the Guaranioccupying an intermediate position Essentially the same results were obtainedwith the principal coordinate and principal components analyses (data not shown)

Average heterozygosities considering the Alu insertions and the blood group plusprotein systems results showed essentially the same patterns with the AcheAcirc havingsomewhat lower values (015 for both sets) while the three other groups are moreuniform (021plusmn023 025plusmn029 respectively) But since the standard errors are highthe diŒerences are statistically non-signiregcant Total variability and the amount of it

that is due to inter-population diŒerences were similar considering the Alu poly-morphisms (026 8) and the blood group plus protein results (023 10)

We are now in a position to answer the two questions posed in the Introduction(a) yes the intertribal patterns of relationship and other aspects of their variationshow excellent congruence in the two sets of systems and (b) due to the markedgenetic peculiarities of the AcheAcirc we cannot decide between the two hypotheses con-cerning their classiregcation but presently the view that they are a diŒerentiatedGuarani group seems more likely The point to be emphasized however is theirdistinctiveness in relation to the other Amerindians in general Unpublished resultswe have in other genetic systems are pointing in the same direction and a previousanalysis made by Salzano and Callegari-Jacques (1988) showed that in a dendrogramobtained comparing 58 South American Indian groups uniformly studied for seven


Figure 2 Dendrograms obtained with the neighbour-joining method using the genetic distances dis-played in table 5 and mid-point rooting (a) Protein plus blood group systems (b) Alu elements

Table 5 Matrix of genetic distances Above diagonal blood group plus protein systems below diagonalAlu insertion frequencies

AcheAcirc Caingang Guarani Xavante

AcheAcirc ETH 00410 00314 00400Caingang 00499 ETH 00091 00138Guarani 00300 00214 ETH 00072Xavante 00564 00211 00148 ETH

genetic systems (MNSs P Rh DuŒy Kidd Diego and haptoglobin) the AcheAcirc

clearly diŒerentiated from all of them with a single exception They clustered withthe ParakanaAuml who live 2500 km away in the Amazon region but who also speak aTupiplusmnGuarani language and have like the AcheAcirc light skin

AcknowledgementsThanks are due to the FundacEuml aAuml o Nacional do Indio (FUNAI) for permission to

study the Indians and help in the regeld and to the FundacioAcirc n Bertoni for logisticassistance The Indian leaders and the subjects of the investigation were appropri-ately informed about the aims of the study and gave their approval which is grate-fully acknowledged The investigation was also approved by the Brazilian NationalEthics Commission (CONEP Resolution no 12398) Financial support was pro-vided by Programa de Apoio a NuAcirc cleos de ExceleAtilde ncia (PRONEX Brazil)Financiadora de Estudos e Projetos (FINEP Brazil) Conselho Nacional deDesenvolvimento CientotildeAcirc regco e TecnoloAcirc gico (CNPq Brazil) FundacEuml aAuml o de AmparoaAacute Pesquisa do Estado do Rio Grande do Sul (FAPERGS Brazil) Louisiana Boardof Regents Millennium Trust Health Excellence Fund HEF (2000-05)-05 and HEF(2000-05)-01 National Science Foundation (NSF USA) and National Institutes ofHealth (NIH USA)

AppendixBlood group and protein genetic systems allele or haplotype frequencies observed

among the Caingang Guarani and Xavante

Allele orhaplotype Caingang Guarani Xavante

System frequency hellipn ˆ 35daggery hellipn ˆ 99daggery hellipn ˆ 85daggery

ABO ABOO 100 100 100MNSs LMS 036 029 029

LMs 036 035 043LNS 012 009 016LNs 016 027 012

P P1 039 059 052Rh RHCDE 001 001 000

RHCDe 044 060 056RHcDE 039 022 016RHcDe 008 014 028RHcde 008 003 000

Kell KELLk 100 100 100DuŒy FYA 046 058 045Haemoglobin HBA 100 100 100Glucose-6-phosphate-dehydrogenase G6PDB 100 100 100Phosphogluconate dehydrogenase PGDA 100 100 100Phosphoglucomutase 1 PGM11 086 082 085Phosphoglucomutase 2 PGM21 100 100 100Adenylate kinase AK1 100 100 100Acid phosphatase ACPB 089 088 078Esterase D ESD1 089 049 063Glyoxalase 1 GLO1 025 018 022Haptoglobin HP1 069 059 055Transferrin TFC 100 100 085Ceruloplasmin CPB 100 100 100GC GC1 034 042 074Albumin ALBA 100 100 100

Based on data reported as follows Caingang Salzano and Shresbquoer (1966) Salzano Callegari-JacquesFranco et al (1980) Guarani Salzano et al (1997a) Xavante Salzano et al (1997b)y Modal number of individuals studied


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Address for correspondence Prof Francisco M Salzano Departamento de GeneAcirc tica Instituto deBiocieAtilde ncias Universidade Federal do Rio Grande do Sul Caixa Postal 15 053 91 501-970 Porto AlegreRS Brazil email franciscosalzanoufrgsbr

Zusammenfassung Hintergrund Sind die Verwandtschaftsbeziehungen zwischen Populationen dieanhand der DNA ermittelt wurden die gleichen wie die welche anhand der Blutgruppe und derProteinmarker gefunden wurden Zeigen sich Unterschiede die darauf hinweisen dass die Faktorenwelche die genetische VariabilitaEgrave t auf diesen zwei Niveaus der Analyse beeinmacrussen verschieden sindKoEgrave nnen diese Marker bei der biologischen Klassiregkation der AcheAcirc einem paraguayischen Stamm der erstvor kurzem staEgrave ndigen Kontakte mit Nicht-Indianern aufnahm genutzt werdenMaterial und Methoden Zur KlaEgrave rung dieser Fragen wurden 193 Individuen von vier AmerindianischenStaEgrave mmen in Bezug auf 12 Alu Polymorphismen typisiert (fuEgrave nf davon wurden in diesen Populationen bishernoch nie untersucht) waEgrave hrend 22 Blutgruppen und Proteinsysteme der AcheAcirc untersucht wurden DieseDaten wurden dann zu jenen Daten (Blutgruppen und Proteine) in Beziehung gesetzt welche vorher fuEgrave rdie drei anderen Populationen vorhandenen waren DNA Extraktion und Ampliregkation sowie andereLaboranalysen wurden in unserem Labor mit aktuell gebraEgrave uchlichen Standardmethoden durchgefuEgrave hrtDie genetischen Verwandtschaften wurden mittels DA plusmnDistanz ermittelt und die StammbaEgrave ume durch dieneighbour-joining Methode konstruiert welche von M Nei et al entwickelt wurde Die ZuverlaEgrave ssigkeitder StammbaEgrave ume wurde durch Bandenreplikationen gepruEgrave ft Andere VariabilitaEgrave ten bei den Populationenwurden auch mit Neirsquos Methoden bestimmtErgebnisse Alu Polymorphismen wurden in allen Populationen und fuEgrave r die meisten Loci beobachtet inden sieben Systemen in denen wir unsere Resultate mit denen anderer Amerindianischer Gruppen ver-gleichen konnten war die UEgrave bereinstimmung zufriedenstellend UngewoEgrave hnliche Befunde bei derBlutgruppe und den Proteinsystemen der AcheAcirc waren eine sehr niedrig (5) HP1 Frequenz und dasVorhandensein eines CW PhaEgrave notypes bei der Rh Blutgruppe Die Verwandschaftsbeziehungen zwischenden StaEgrave mmen und andere Aspekte ihrer VariabiltaEgrave t waren bei den zwei Systemen (Alu Blutgruppe undProtein) bemerkenswert uEgrave bereinstimmendSchlussfolgerungen Die Antwort auf die erste der aufgeworfenen Fragen ist positiv Jedoch laEgrave sst sich dasProblem ob die AcheAcirc von einer GeAtilde Gruppe abstammen welche der Guarani Besiedlung von Paraguayvoranging oder eine unterschiedliche Guarani Gruppe darstellen nicht mit den verfuEgrave gbaren genetischenInformationen loEgrave sen Die zweite Hypothese scheint zur Zeit wahrscheinlicher aber ein Punkt der hervor-gehoben werden muss ist die auŒallende genetische Besonderheit der AcheAcirc verglichen mit anderenAmerindians

ReAcirc sumeAcirc ArrieAacutere-plan Est-ce que les relations entre populations observeAcirc es au moyen drsquoADN ou degroupes sanguins plus marqueurs proteAcirc iques demeurent identiques ou bien reAcirc veAacute lent-elles des dispositionsdiŒeAcirc rentes qui indiqueraient que les facteurs qui inmacruencent la variation geAcirc neAcirc tique aAacute ces deux niveaux sontdiŒeAcirc rents Ces marqueurs peuvent-ils aider aAacute reAcirc pertorier les AcheAcirc une ethnie paraguayenne ni nrsquoa quereAcirc cemment eAcirc tabli des contacts plus freAcirc quents avec des non indiensSujets et meAcircthodes Aregn de reAcirc pondre aAacute ces questions on a typeAcirc 193 individus de 4 ethnies ameAcirc rindiennespour 12 polymorphismes Alu (dont 5 nrsquoont jamais eAcirc teAcirc eAcirc tudieAcirc s dans ces populations) tandis que 22 groupessanguins et seAcirc riques ont eAcirc teAcirc eAcirc tudieAcirc s chez les AcheAcirc Ces donneAcirc es ont eAcirc teAcirc ensuite inteAcirc greAcirc es aAacute celles qui eAcirc taientdeAcirc jaAacute disponibles (groupe sanguins et seAcirc riques) pour les trois autres populations Lrsquoextraction et lrsquoamplireg-cation de lrsquoADN ainsi que drsquoautres proceAcirc dures de laboratoire ont eAcirc teAcirc reAcirc aliseAcirc es par les meAcirc thodes standardhabituellement utiliseAcirc es par notre laboratoire Les associations geAcirc neAcirc tiques ont eAcirc teAcirc obtenues au moyen de ladistance DA et les dendrogrammes construits par la meAcirc thode de regroupement de voisinage eAcirc tablie par MNei et collaborateurs La validiteAcirc des dendrogrammes a eAcirc teAcirc testeAcirc e par reAcirc plications croiseAcirc es Drsquoautresvaleurs de variabiliteAcirc populationnelle ont eAcirc galement eAcirc teAcirc deAcirc termineAcirc es par les meAcirc thodes de NeiReAcircsultats Le polymorphisme Alu est observeAcirc dans toutes les populations et pour la plupart des loci Danssept systeAacute mes aAacute partir desquels nous avons pu comparer nos reAcirc sultats avec ceux drsquoautres groupes ameAcirc r-indiens lrsquoagreAcirc ment a eAcirc teAcirc satisfaisant Des reAcirc sultats inhabituels sur les groupes sanguins et seAcirc riques desAcheAcirc portaient sur une freAcirc quence treAacute s basse (5) de HP1 et sur la preAcirc sence du pheAcirc notype CW dans lesysteAacute me Rh Les scheAcirc mas intertribaux drsquoassociation et drsquoautres aspects de leur variation eAcirc taient remar-quablement congruents dans les deux seAcirc ries de systeAacute mes (Alugroupes sanguins et seAcirc riques)Conclusions La reAcirc ponse aAacute la premieAacute re question est a rmative Par contre le probleAacute me de savoir si les AcheAcircsont des deAcirc riveAcirc s drsquoun groupe GeAtilde qui a preAcirc ceAcirc deAcirc la colonisation du Paraguay par les Guarani ou sontseulement un groupe guarani diŒeAcirc rencieAcirc nrsquoa pas pu eAtilde tre reAcirc solu par les donneAcirc es geAcirc neAcirc tiques disponiblesLa seconde hypotheAacute se paraotildeAtilde t aAacute preAcirc sent plus probable mais le point qui doit eAtilde tre eAcirc clairci est celui de lafrappante singulariteAcirc des AcheAcirc par rapport aux autres ameAcirc rindiens


postulated to be retrotransposons that have been inserted into the human genomevia a single-stranded RNA intermediate generated by RNA polymerase III tran-scription (Weiner Deininger and Efstratiadis 1986)

There are approximately 2000 Ya58 and 2000 Yb8 insertional elements randomlydispersed throughout the human genome Many of these remacrect recent retropositionevents that have not yet been regxed within the human species (Batzer and Deiniger1991 Batzer Stoneking Alegria-Hartmann et al 1994 Batzer Rubin Hellmann-Blumberg et al 1995 Novick Novick Yunis et al 1995 Arcot De Angelis Sherryet al 1997 Stoneking Fontius CliŒord et al 1997 Novick Novick Yunis et al1998) Due to changes during the evolution of the source genes there are at least 12major Alu subfamilies which may be classireged as old intermediate or young Thetwo oldest subfamilies Jo and Jb arose from independent retroposition eventsinvolving a single ancestral source gene that occurred early in primate evolution(Kapitonov and Jurka 1996) The polymorphic Alu elements may serve as markers toelucidate various aspects of human evolution There are many advantages for theiruse as genetic markers in human populations It is highly improbable that a speciregcAlu sequence has retroposed into a particular site more than once in human evolu-tion and recent insertions represent stable polymorphisms that are rarely lost with-out leaving a trace Because of these two properties identity by descent andevidential stability due to incomplete deletion Alu insertions are literally molecularfossils Another attractive feature of Alu-based population studies is that the ances-tral condition is deregned by the lack of an Alu element Knowledge of the originalcharacter state is useful in rooting phylogenies A further advantage of Alu geneticmarkers is that Alu genotypes involving individual or multiple polymorphic loci canbe determined by a rapid PCR-based assay

For many decades there have been extensive investigations on the biologicalcharacteristics of South Amerindians (Salzano and Callegari-Jacques 1988)Although in recent years the studies have included investigations at the DNAlevel most of the data have been gathered from morphological characteristics ortraditional genetic markers such as blood groups and proteins Despite the consid-erable eŒorts employed the patterns of genetic variability among South Americannative populations are not yet su ciently clear (ClariaAcirc Demarchi Moreno Azoreroet al 1998)

The present study furnishes new information on 12 Alu insertion polymorphismsfor four Amerindian groups as well as blood group and protein genetic data for oneof them (the AcheAcirc of Paraguay) Previous results concerning these latter systemswere compiled for the three other populations and the following questions wereaddressed (a) Are the patterns of relationships involving these four groups thesame considering the two sets of characteristics and (b) Can they provide informa-tion about how the AcheAcirc should be classireged There are competing hypotheses in thisregard some scholars considering them a GeAtilde group that preceded the Guaranicolonization of Paraguay while others contend that they are just a diŒerentiatedGuarani group

2 Subjects and methodsGeneral information about the four populations studied is presented in table 1

and reggure 1 They live in the centralplusmnsouthern part of the continent in Paraguay(AcheAcirc ) and Brazil (the other three) speak languages classireged in the Tupi and GeAtilde plusmnKaingan linguistic groups and the degree of non-Indian socio-cultural inmacruence





























Population
































Table

2P

rese

nce

of

Alu

inse

rtio

ns

am

ong

mem

ber

sof

four

South

Am

eric

an

Ind

ian

popu

lati

on

s

Ach

eAcircC

aingan

gG

uar

ani

Xav

ante

Alu

No

of

Allel

eN

o

of

Allel

eN

o

of

Allel

eN

o

of

Allel

ein

sert

ion

Gen

oty

pe

indiv

iduals

freq

uen

cies

indiv

iduals

freq

uen

cies

indiv

iduals

freq

uen

cies

indiv

idu

als

freq

uen

cies

FX

IIIB

DaggerDagger

45

0782

320

872

2709

35

3010

00

Dagger21

114

05

00

0H

Weq

p

02

96

0001

p10

00

00

00

p10

0000

00

NA

475

DaggerDagger

74

1000

401

000

2710

00

2910

00

Dagger0

00

00

00

0H

Weq

N

AN

AN

AN

A

MA

BD

1Dagger

Dagger10

0581

320

706

903

94

1005

32

Dagger16

18

135

1316

8H

Weq

p

10

00

0000

p00

00

00

00

p00

0900

00

p

04

7100

01

323

DaggerDagger

61

0908

90

420

2008

00

1707

12

Dagger7

248

133

172

3H

Weq

p

00

08

0000

p10

00

00

00

p03

0800

01

p

10

0000

00

A25

DaggerDagger

00

013

00

037

000

97

102

34

Dagger2

36

1373

3825

18H

Weq

p

10

00

0000

p10

00

00

00

p10

0000

00

p

06

5200

01

TP

A25

DaggerDagger

34

0866

170

675

1707

10

604

17

Dagger3

2010

134

34

11H

Weq

p

00

01

0000

p04

87

00

02

p02

1900

01

p

07

0500

01


9Dagger

Dagger29

09

68

31

08

54

16

06

5215

0717

Dagger2

811

13

02

62

HW

eq

p

1000

00

00p

0181

0001

p

0137

0001

p

10

00

0000

46

5Dagger

Dagger0

00

00

201

59

000

210

0078

Dagger0

91

531

30

23

27

HW

eq

NA

p

0240

0001

p

1000

0000

p

10

00

0000

AP

ODagger

Dagger36

10

00

37

09

63

32

09

4133

1000

Dagger0

30

00

02

0H

Weq

N

Ap

1000

0000

p

0001

0000

NA

43

2Dagger

Dagger7

01

98

202

50

101

302

0242

Dagger11

20

511

45

26

21

18

HW

eq

p

0001

00

00p

0700

0001

p

0356

0001

p

10

00

0000

PV

92

DaggerDagger

51

08

55

28

07

93

20

07

8323

0813

Dagger16

97

62

43

3H

Weq

p

0619

00

01p

0449

0001

p

0100

0001

p

00

47

0001

AC

EDagger

Dagger76

10

00

17

05

43

25

08

2915

0683

Dagger0

16

811

013

24

HW

eq

NA

p

0043

0001

p

0235

0001

p

04

17

0002

T

he

pre

sen

cean

dabse

nce

of

the

Alu

repea

tar

eden

ote

dby

Daggerand

re

spec

tivel

y

HW

eq

Exac

tte

stfo

rH

ardyplusmnW

einber

geq

uilib

rium

(pro

babilit

yva

lue

plu

sor

min

us

its

standard

erro

r)

NA

N

ot

applica

ble





































P P1 97 P1 0858 NAP2 2




p lt 005









2-2 5 p gt 070

























ABO ABOO 100 100 100MNSs LMS 036 029 029

LMs 036 035 043LNS 012 009 016LNs 016 027 012

P P1 039 059 052Rh RHCDE 001 001 000


























































































Population
































Table

2P

rese

nce

of

Alu

inse

rtio

ns

am

ong

mem

ber

sof

four

South

Am

eric

an

Ind

ian

popu

lati

on

s

Ach

eAcircC

aingan

gG

uar

ani

Xav

ante

Alu

No

of

Allel

eN

o

of

Allel

eN

o

of

Allel

eN

o

of

Allel

ein

sert

ion

Gen

oty

pe

indiv

iduals

freq

uen

cies

indiv

iduals

freq

uen

cies

indiv

iduals

freq

uen

cies

indiv

idu

als

freq

uen

cies

FX

IIIB

DaggerDagger

45

0782

320

872

2709

35

3010

00

Dagger21

114

05

00

0H

Weq

p

02

96

0001

p10

00

00

00

p10

0000

00

NA

475

DaggerDagger

74

1000

401

000

2710

00

2910

00

Dagger0

00

00

00

0H

Weq

N

AN

AN

AN

A

MA

BD

1Dagger

Dagger10

0581

320

706

903

94

1005

32

Dagger16

18

135

1316

8H

Weq

p

10

00

0000

p00

00

00

00

p00

0900

00

p

04

7100

01

323

DaggerDagger

61

0908

90

420

2008

00

1707

12

Dagger7

248

133

172

3H

Weq

p

00

08

0000

p10

00

00

00

p03

0800

01

p

10

0000

00

A25

DaggerDagger

00

013

00

037

000

97

102

34

Dagger2

36

1373

3825

18H

Weq

p

10

00

0000

p10

00

00

00

p10

0000

00

p

06

5200

01

TP

A25

DaggerDagger

34

0866

170

675

1707

10

604

17

Dagger3

2010

134

34

11H

Weq

p

00

01

0000

p04

87

00

02

p02

1900

01

p

07

0500

01


9Dagger

Dagger29

09

68

31

08

54

16

06

5215

0717

Dagger2

811

13

02

62

HW

eq

p

1000

00

00p

0181

0001

p

0137

0001

p

10

00

0000

46

5Dagger

Dagger0

00

00

201

59

000

210

0078

Dagger0

91

531

30

23

27

HW

eq

NA

p

0240

0001

p

1000

0000

p

10

00

0000

AP

ODagger

Dagger36

10

00

37

09

63

32

09

4133

1000

Dagger0

30

00

02

0H

Weq

N

Ap

1000

0000

p

0001

0000

NA

43

2Dagger

Dagger7

01

98

202

50

101

302

0242

Dagger11

20

511

45

26

21

18

HW

eq

p

0001

00

00p

0700

0001

p

0356

0001

p

10

00

0000

PV

92

DaggerDagger

51

08

55

28

07

93

20

07

8323

0813

Dagger16

97

62

43

3H

Weq

p

0619

00

01p

0449

0001

p

0100

0001

p

00

47

0001

AC

EDagger

Dagger76

10

00

17

05

43

25

08

2915

0683

Dagger0

16

811

013

24

HW

eq

NA

p

0043

0001

p

0235

0001

p

04

17

0002

T

he

pre

sen

cean

dabse

nce

of

the

Alu

repea

tar

eden

ote

dby

Daggerand

re

spec

tivel

y

HW

eq

Exac

tte

stfo

rH

ardyplusmnW

einber

geq

uilib

rium

(pro

babilit

yva

lue

plu

sor

min

us

its

standard

erro

r)

NA

N

ot

applica

ble





































P P1 97 P1 0858 NAP2 2




p lt 005









2-2 5 p gt 070

























ABO ABOO 100 100 100MNSs LMS 036 029 029

LMs 036 035 043LNS 012 009 016LNs 016 027 012

P P1 039 059 052Rh RHCDE 001 001 000




















































































Table

2P

rese

nce

of

Alu

inse

rtio

ns

am

ong

mem

ber

sof

four

South

Am

eric

an

Ind

ian

popu

lati

on

s

Ach

eAcircC

aingan

gG

uar

ani

Xav

ante

Alu

No

of

Allel

eN

o

of

Allel

eN

o

of

Allel

eN

o

of

Allel

ein

sert

ion

Gen

oty

pe

indiv

iduals

freq

uen

cies

indiv

iduals

freq

uen

cies

indiv

iduals

freq

uen

cies

indiv

idu

als

freq

uen

cies

FX

IIIB

DaggerDagger

45

0782

320

872

2709

35

3010

00

Dagger21

114

05

00

0H

Weq

p

02

96

0001

p10

00

00

00

p10

0000

00

NA

475

DaggerDagger

74

1000

401

000

2710

00

2910

00

Dagger0

00

00

00

0H

Weq

N

AN

AN

AN

A

MA

BD

1Dagger

Dagger10

0581

320

706

903

94

1005

32

Dagger16

18

135

1316

8H

Weq

p

10

00

0000

p00

00

00

00

p00

0900

00

p

04

7100

01

323

DaggerDagger

61

0908

90

420

2008

00

1707

12

Dagger7

248

133

172

3H

Weq

p

00

08

0000

p10

00

00

00

p03

0800

01

p

10

0000

00

A25

DaggerDagger

00

013

00

037

000

97

102

34

Dagger2

36

1373

3825

18H

Weq

p

10

00

0000

p10

00

00

00

p10

0000

00

p

06

5200

01

TP

A25

DaggerDagger

34

0866

170

675

1707

10

604

17

Dagger3

2010

134

34

11H

Weq

p

00

01

0000

p04

87

00

02

p02

1900

01

p

07

0500

01


9Dagger

Dagger29

09

68

31

08

54

16

06

5215

0717

Dagger2

811

13

02

62

HW

eq

p

1000

00

00p

0181

0001

p

0137

0001

p

10

00

0000

46

5Dagger

Dagger0

00

00

201

59

000

210

0078

Dagger0

91

531

30

23

27

HW

eq

NA

p

0240

0001

p

1000

0000

p

10

00

0000

AP

ODagger

Dagger36

10

00

37

09

63

32

09

4133

1000

Dagger0

30

00

02

0H

Weq

N

Ap

1000

0000

p

0001

0000

NA

43

2Dagger

Dagger7

01

98

202

50

101

302

0242

Dagger11

20

511

45

26

21

18

HW

eq

p

0001

00

00p

0700

0001

p

0356

0001

p

10

00

0000

PV

92

DaggerDagger

51

08

55

28

07

93

20

07

8323

0813

Dagger16

97

62

43

3H

Weq

p

0619

00

01p

0449

0001

p

0100

0001

p

00

47

0001

AC

EDagger

Dagger76

10

00

17

05

43

25

08

2915

0683

Dagger0

16

811

013

24

HW

eq

NA

p

0043

0001

p

0235

0001

p

04

17

0002

T

he

pre

sen

cean

dabse

nce

of

the

Alu

repea

tar

eden

ote

dby

Daggerand

re

spec

tivel

y

HW

eq

Exac

tte

stfo

rH

ardyplusmnW

einber

geq

uilib

rium

(pro

babilit

yva

lue

plu

sor

min

us

its

standard

erro

r)

NA

N

ot

applica

ble





































P P1 97 P1 0858 NAP2 2




p lt 005









2-2 5 p gt 070

























ABO ABOO 100 100 100MNSs LMS 036 029 029

LMs 036 035 043LNS 012 009 016LNs 016 027 012

P P1 039 059 052Rh RHCDE 001 001 000
































































9Dagger

Dagger29

09

68

31

08

54

16

06

5215

0717

Dagger2

811

13

02

62

HW

eq

p

1000

00

00p

0181

0001

p

0137

0001

p

10

00

0000

46

5Dagger

Dagger0

00

00

201

59

000

210

0078

Dagger0

91

531

30

23

27

HW

eq

NA

p

0240

0001

p

1000

0000

p

10

00

0000

AP

ODagger

Dagger36

10

00

37

09

63

32

09

4133

1000

Dagger0

30

00

02

0H

Weq

N

Ap

1000

0000

p

0001

0000

NA

43

2Dagger

Dagger7

01

98

202

50

101

302

0242

Dagger11

20

511

45

26

21

18

HW

eq

p

0001

00

00p

0700

0001

p

0356

0001

p

10

00

0000

PV

92

DaggerDagger

51

08

55

28

07

93

20

07

8323

0813

Dagger16

97

62

43

3H

Weq

p

0619

00

01p

0449

0001

p

0100

0001

p

00

47

0001

AC

EDagger

Dagger76

10

00

17

05

43

25

08

2915

0683

Dagger0

16

811

013

24

HW

eq

NA

p

0043

0001

p

0235

0001

p

04

17

0002

T

he

pre

sen

cean

dabse

nce

of

the

Alu

repea

tar

eden

ote

dby

Daggerand

re

spec

tivel

y

HW

eq

Exac

tte

stfo

rH

ardyplusmnW

einber

geq

uilib

rium

(pro

babilit

yva

lue

plu

sor

min

us

its

standard

erro

r)

NA

N

ot

applica

ble





































P P1 97 P1 0858 NAP2 2




p lt 005









2-2 5 p gt 070

























ABO ABOO 100 100 100MNSs LMS 036 029 029

LMs 036 035 043LNS 012 009 016LNs 016 027 012

P P1 039 059 052Rh RHCDE 001 001 000



































































































P P1 97 P1 0858 NAP2 2




p lt 005









2-2 5 p gt 070

























ABO ABOO 100 100 100MNSs LMS 036 029 029

LMs 036 035 043LNS 012 009 016LNs 016 027 012

P P1 039 059 052Rh RHCDE 001 001 000
















































































ABO ABOO 100 100 100MNSs LMS 036 029 029

LMs 036 035 043LNS 012 009 016LNs 016 027 012

P P1 039 059 052Rh RHCDE 001 001 000































































alu insertions versus blood group plus protein genetic variability in four amerindian populations

Documents