trehalose and carbohydrate metabolism of l. bicolor a. deveau p. frey-klett, j.garbaye, f. martin et...
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TrehaloseTrehaloseand carbohydrate and carbohydrate
metabolism of metabolism of L. bicolorL. bicolor
A. Deveau
P. Frey-Klett, J.Garbaye, F. Martin
et al.
UMR INRA-UHP 1136 Interactions arbres micro-organismes UFR 110
© INRA
Carbohydrate metabolismCarbohydrate metabolism
KEGG Pathway database
• glycolysis and TCA cycle• pentose phosphate pathway• malate synthesis• trehalose, glycogen metabolism
Complete pathways:
Uncomplete pathways:
• mannose (mannose-1-phosphate
guanylyltransferase …)
• mannitol
F-6-P
Mannose-6-P
Mannitol
Mannitol-1-PF-6-P
Phosphomannose isomerase
Mannose-6-P reductase
Mannitol-1-P-phosphataseMannitol-1-P-phosphatase
Mannitol dehydrogenase
http://www.genome.jp/kegg/pathway/map/map01110.html
Trehalose and
Glycogen metbolism
The trehalose in The trehalose in ectomycorrhizal fungi ectomycorrhizal fungi
glucose
fructose
glucose
fructose
sucrosesucrose
fructose
glucose
+
F6P
glycogen
trehalose
mannitol
glycolysis
plant cortical cellplant cortical cellfungal hyphaefungal hyphae
apoplast
G6P
CH2OHCH2OH*
• Main roles:Main roles:
• Storage sugar – supplies of endogenous carbon
• Stress protectant by stabilization of cellular membranes and proteins (freezing, dessication, growth restriction)
• A signal molecule
photosynthesis
transport
The trehalose in The trehalose in ectomycorrhizal fungi ectomycorrhizal fungi
•Involved in interactions between mycorrhization helper bacteria and Laccaria bicolor ?
• Strongly accumulated in Laccaria hyphae
L. Bicolor hyphae
Pseudomonas fluorescens
trehalosetrehalose
Bacterial effect on Bacterial effect on presymbiotic growth presymbiotic growth
and survival and survival of the fungusof the fungus
ChemiotactismChemiotactism
Improvement of Improvement of bacterial survivalbacterial survival
Tsl 1Tsl 1
Trehalose 6-P
TrehaloseUDP-Glucose
Glucose 6-P
Glucose 1-P
Pi
glycogen metabolism
glycolyse
Glucose
PiHxk 1Hxk 1Hxk 2Hxk 2
Ugp 1Ugp 1
PgmPgmTps 1Tps 1 Tps 2Tps 2
Glucose TrehaloseTre 2Tre 2
TP ?TP ?extracellular
intracellular
vacuolarstorage
Glucose
Tre 1Tre 1 TPTP
The putative metabolism of trehalose in The putative metabolism of trehalose in L.bicolorL.bicolor
TPS3TPS3
Tsl 1Tsl 1
Trehalose 6-P
TrehaloseUDP-Glucose
Glucose 6-P
Glucose 1-P
Pi
glycogen metabolism
glycolyse
Glucose
PiHxk 1Hxk 1Hxk 2Hxk 2
Ugp 1Ugp 1
PgmPgmTps 1Tps 1 Tps 2Tps 2
Glucose TrehaloseTre 2Tre 2
TP ?TP ?extracellular
intracellular
vacuolarstorage
Glucose
Tre 1Tre 1 TPTP
Specificities of the Specificities of the L. bicolor L. bicolor metabolismmetabolism
Trehalose phosphorylase
C. neoformansS. cerevisiae
trehalose transporter
Genes structure :Genes structure :
Feature
Gene length (bp)
Coding size (aa)
Exons number per gene
Mean exon size (bp)
Mean intron size (bp)
2076
Glk1
Notes :Notes :
• The genes are scattered on 9 different scaffolds. TPS1 and Tre2 on the 11th scaffold, separated by 200 000 bp.
• Two categories of gene structure: high similarity with Phanerochaete gene structure vs low similarity
• Variable intron size
Glk2 UGP1 PGM TPS1 TPS2 TSL1 Tre1 Tre 2 TP
2276 2040 1968 1836 3749 3214 3063 2747 2851
479 484 523 570 511 957 999 727 756 738
13 12 10 5 6 14 9 17 9 13
117
53 53
128 252
60
205
68
256
61
365
65
170
53
157
5273
123
P. chrysosporum
P. chrysosporum
P. chrysosporum
P. chrysosporum
P. chrysosporum
342
69
2406
502
11
132
90
2127
516
12
129
53
1864
566
4
424
55
2685
783
7
56
324
2532
667
8
75
3540
869
9
274
117
1803
526
5
303
56
2425
678
9
367
49
-
-
-
-
-
1849
474
9
176
53
220
Bases composition of « large introns » (> 70 bp)Bases composition of « large introns » (> 70 bp)
•High poly THigh poly T content content•No identityNo identity with with transposonstransposons sequences from Genebank sequences from Genebank
Intron n° 3 of TSL1 (93 bp) GTAAGTTTTTTTTGCTATCTTTTTTTTGTTTTTTTTTGTTTTTGTTTTTGTTTTTTTGTT TGTTTGGGAGATGGGTGCTAATGATGTTTTTAG Intron n° 8 of TSL1 (100 bp) GTTTAGGACCTCCGCTTTTTTTCTCTGCGGACATTGATCGCTGCTCTTTCTTTTATTCTC CGTCTCCCGTTTGTTTTCTTGATGGGCTAACACCTTCAAG
Sequence evolution Sequence evolution
© Broad Institute
• high degree of amino acid identity with other fungi(> 70 %)
Acid trehalase (Tre2)Trehalose synthase 1 (TPS1)Trehalose phosphorylase (TP)
Uridyltransferase (UGP1)Hexokinase 2 (Glk 2)
• low degree of amino acid identity (< 65 %)Neutral trehalase (Tre 1)
Trehalose phosphorylase (TPS2)Trehalose synthase - regulatory sub-unit (TSL1)
Hexokinase 1 (Glk1)
Two groups of protein:
Sequence evolution Sequence evolution
M. griseusG. zeaeN. crassaA. nigraA. fumigatusA. nidulansC. neoformensL. BicolorA. muscaria
Concensus
M. griseusG. zeaeN. crassaA. nigraA. fumigatusA. nidulansC. neoformensL. BicolorA. muscaria
Concensus
M. griseusG. zeaeN. crassaA. nigraA. fumigatusA. nidulansC. neoformensL. BicolorA. muscaria
Concensus
M. griseusG. zeaeN. crassaA. nigraA. fumigatusA. nidulansC. neoformensL. BicolorA. muscaria
Concensus
Trehalose synthase 1 (TPS1)Trehalose synthase 1 (TPS1)
High level of aminoacid sequence conservation (> 70%)High level of aminoacid sequence conservation (> 70%)
Sequence evolution Sequence evolution
Hexokinase 1 (GLK1)Hexokinase 1 (GLK1)
Lower level of aminoacid sequence conservation (< 65%)Lower level of aminoacid sequence conservation (< 65%)
N. crassaU. triticiM. griseusA. fumigatusL. bicolorP. involutusC. neoformensT. reesei
consensus
N. crassaU. tritciM. griseusA. fumigatusL. bicolorP. involutusC. neoformensT. reesei
consensus
N. crassaU. triticiM. griseusA. fumigatusL. bicolorP. involutusC. neoformensT. reesei
consensus
N. crassaU. triticiM. griseusA. fumigatusL. bicolorP. involutusC. neoformensT. reesei
consensus
Sequence evolution Sequence evolution
82 %
72 %
72 %
62 %
63 %65 %
66 %
74 %
Trehalose synthase 1 (TPS1)
• High degree of identity (>70 %)
Acid trehalase (Tre2)
Trehalose synthase 1 (TPS1)
Trehalose phosphorylase (TP)
Uridyltransferase (UGP1)
Hexokinase 2 (Glk 2)
• Lower degree of identity (< 65%)
Hexokinase 1
61 %
58 %
53 %
42 %
42 %42 %
41 %
36 %
38 %
Neutral trehalase (Tre 1)
Trehalose phosphorylase (TPS2)
Trehalose synthase - regulatory sub-unit (TSL1)
Hexokinase 1 (Glk1)
Ascomycetes
Ascomycetes
Basidiomycetes
Basidiomycetes
• Carbohydrates metabolism:
No main sugar biosynthesis pathway
missing
• Trehalose
• metabolic specifitiesmetabolic specifities -> two
ways of synthesis (trehalose trehalose
phosphorylasephosphorylase)
• presence of large intronslarge introns with
polyT repeatspolyT repeats
• variable degree of variable degree of
conservation of aminoacid conservation of aminoacid
sequencesequence
CH2OHCH2OH*
SummarySummary
© INRA
The bi-phospho-fructo aldolase, a duplicated gene ?The bi-phospho-fructo aldolase, a duplicated gene ?
scaffold 25 scaffold 12
• nucleotidique sequence: 77 % of identity
amino acid sequence: 90 % of identity • Exactly same exon/intron structure and sizes •No identity between UTR sequences and surounding sequences• EST for one sequence: scaffold 25