第六章 基因表达的调控 本章,我们讨论 4 个问题: 1....
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第六章 基因表达的调控 本章,我们讨论 4 个问题: 1. 什么是基因表达的调控? 2. 原核生物基因表达调控是如何进行的? 3. 真核生物基因表达调控是如何进行的? 4. 基因表达调控与医学研究. 概 述 一、基因( gene) ( 一 ) 概念 从化学上来说指的是一段 DNA 或 RNA (病毒)顺序,该顺序可以产生或影响某种表型,可以由于突变生成等位基因变异体。 从遗传学上来说代表一个遗传单位、 1 个功能单位, 1 个交换单位和 1 个突变单位。 - PowerPoint PPT PresentationTRANSCRIPT
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41.2.3.4.
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gene)() DNARNA 111 RNA7P27
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1. PhenotypegenetypeLeu+Leu-Leu+Leu- 2.allele 1222DNA
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3. 4. 5. 6.
structural gene mRNArRNAtRNADNA
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genome 11112111 3
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1. RNA polRNARNADNARNA 2. proteintRNAmRNA 1110
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DNAprotein
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()prokaryote DNAprotein1 eukaryote2
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11
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1.2.RNA pol DNA pol DNA compt 1.2. 3.
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100Britten-Davidsen RNA pol
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1 2
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1house keeping geneconstitute expression 2indutioninducible genes
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60JacobMonodE.coli adaptive enzymeLac operon1 1 11111 1.operon 1mRNACAP
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2. P-O
1(inciduble operon)(induction)(inducer)2(repressible operon)(repression)(corepressor)
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DNADNA H1H2H1H21.h2h12.h2h1rh1h1H13.h2-rh1 DNA4.105111
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DNA h2-rh1H2-H1995bp IRLIRR 995bppalindiomeinvertedrepeat sequenceDNA14bp hin hin995bp995bpPh2h2h2-rh1H2-h1h1995bpPh2h2h2-rh1h1 h1h2
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RNA pol: 1. 2.DNAmRNA 3. mRNA
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1.2.
1.promoter2.sigma DNADNADNARNA pol RNA pol2,
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E.coli40~60bp321 +1O2 RNA pol -10bpATTATAboxpribnow3 -35RNApol1PRPDNA2RNA polP2E.coli42~50 32KD 32 32rpohhipR
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-10-3570
4DNAmRNA5Pheat shock responseE.coil. 32,
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3. negative-actingproreinrepressor1OPOPODNARNA polOmRNARNA pol P2,effector 4. positive prateactivator21CAP* 2ntrc* ntrc
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21inducer2corepressorO*1CAP-catabolic gene activator protein*2Ntrc-Ecolinitrogen metabolismntrB
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5.inversion proteinsite-speciFic recombinotion enzymeDNA6.RNA pol-stringent respsnse RNA polRNA polRNArRNA,tRNA tRNAATPpppGpp(-5)ppGpp-4-RNA pol ppGpp-RNA pol RNA pol rRNAtRNA RNA pol rRNAtRNA
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7.teminator 33212 127-20bp216~24bp1G,C 1GC226~8ATbp 8.rho() 6 1 2 NTPaseNTPRNA pol
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DNARNA pol RNA9.attenuator
RNA polRNARNARNADNADNARNA pol GCRNA polread through
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1. 1 -2~3500010005~10% + 2 6019961975
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2Lactose operon Lac operon
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CAPCAP1122
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G- GGLac operonGLac operonG G G cAmp GcAmpcAmpCAPCAPCAPRNA polP CAPCAMP-CAPGG
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Arac-CParc PBAD-IO1O2-B-LisomeraseA-LkinaseD-L4opimeraseI-
2 1 Ara operonCAPAracAra15EcoliAra3Ara5 2Ara operon
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3 aracaraBAD AraC2CindCrep AraAraCind+AraaraBAD araC AraAraCrep araBADaraC Ara operon Lac operonGcAmp-CAP araCaraBAD Ara operonCindCAMP-CAPCrep Ara
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AraAraCreparaO2IDNAaraBAD AraAra+AraCcAMP-CAPCAParaBADaracaraCCindO1C araBADCindBAD
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1.3
(1) RNA poloperonDNA1mRNA(2)1operon DNAPmRNAPRNA(3)2 RNA pol1operon1operon2operon
1
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2.2+1 2 1 2 33 attenutor,A1 AmRNA31 RNA polRNA pol Trp operon
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(trp operoon)1.Trp operon
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2.1TrpO2Trp operon TrpTrp Trpfecdback inhibition TrpROR12.5KD42044OTrp4O Ptrp-10-35-10TrpORNA pol
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3ATrp operon Trp operon1/70Lac operon1/1000Trp operonLac operonTrp aL
aLE30~60bp4GC8UTrp3,42UGGTrp6700bp
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aL 2UGG 4122.33.4GC1.2>2.3>3.4 348U comptaLRNA polaL+90aL TrpTrp-tRNA121.2,2.33.4RNA pol
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TrpTrp-tRNA12Trp2334RNAEDCBA aL TrpmRNAEDCBATrp Trp122
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compt, SD 1.SDSD Sequence,SDs mRNAAuG3~10bp1RbSSD 1SDsAGGAGG 2mRNAAuG4~13bpAuG4~13bp 3A16SRNAUmRNA 411AuGSDSDs
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2.SD 1SDsAuG IL-2PlacSDsAuG 7IL-28IL-2500 2SDs Plac31:0.5:0.2 SDs tRNA
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3.mRNASD SDsmRNA :1omrc 23srRNA((erythromycin methylase))23srRNA2057-2060GAAGA,1amrcTrp operon
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2mRNA
mRNA1.2,2.3,3.4,4.3SD1SD2
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423srRNA1234SD223srRNA1.22.33.4SD223srRNA
23srRNASD2
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mRNA 1.3mRNA mRNAmRNAmRNAmRNA 2.mRNA mRNA 1mRNA 2mRNAmRNARNase3RNasemRNA RNasemRNA 3mRNA2~3minNTPmRNAmRNA
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mRNApr.pr 1. 11pr.mRNA pr.rRNAprEcolipr553421 250pr.pr. 3pr.mRNApr. 41operonmRNA2pr1
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2.RF21 3UAGUGAUAA 1tRNApr.pr. 3 RF1 UAAUAG RF2 UAAUGA RF3 RFRF2 1eIF
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2RF2mRNA reading-framemRNA31Pr.311AuGAuG CCA AAA uuu cccAuG/GCA/AAA/UUU/CCCA/UGG/CAA/AAU/UUC/CC open reading-frameDNARNA 3 Pr.11 Pr mRNA53NC
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mRNA 5 AUGGGG UAU CUU UGAC UAC GAC 3 NH2 Gly Tyr Leu Asp --Tyr -- Asp---COOH 25 315(AA) RF2340Pr.125AAAUG315AA+11 RF221UU26AAASP2RF2UGA25AA 15RF2mRNA 1523 24 25 26 27 28
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RF2 RF2A25UGARF2 RF2 RF2mRNA25AA UGA RF21U 25AA UAG UAG1RF1RF2 25
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RNA1.RNAE.coliPr.Pr.
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1omPR Pr. omPF omPC2OmPR Pr. Ompc ompc mRNA ompc Pr. micF3micFompF mRNAL44SDsAUGompF mRNA42Pr.ompF.ompc5RNAmRNAmRNAmRNAmRNARNA,micRNAmRNA-interfering complementary RNA
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2.RNATn101Tn10terr112Tn10
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3 Tn10pinRNAPout23540bp2RNA 2RNA RNA-inRNA-outRNA-outRNA-in8Tn10 TnTnTn10RNA
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DNA
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DNA1. Rbc 2.gene amplification
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chanrRNArDNA40002000000/500rRNAPr.
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3.gene rearrangement 1 Pr.IgB Ig22DNA1064.gene modificationDNA DNA/DNA/
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/121DNA DNA25CGPr3DNADNaseI
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5. 1 166168bpDNA2Pr.Pr.+RNA8000~10000chromosomechromatin DNA1~15% 2Pr. Pr.histone5510H42AAPr.DNA8H1
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Pr.Pr.NSerArgPrDNA3Pr.non-histon 300~600high mobility group,HMGH1H5DNA
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RNA pol TATAboxconsonsus sequence25~30bpT82A97T93A85AorT100A83AorT83Pr.P
RNA pol-1 12 PDNARNA pol DNA 2 2 ,RNA pol 3I181285.8rRNAsrRNA mRNAPr.-DNATF DTATA+TATA2TFRNA polDNARNA pol P
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1.TFDTATAboxPr.-DNA2.RNA polTF D-DNA3.RNA pol 3
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1.trans-acting factor DNAPr.Pr. DNA8~15DNA12.3 13 DNAPr. 2 3++RNA pol+cis-acting elementDNA
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3.1DNADNA-binding domain 1)zine finger motif DNA2His2cys4cys1Zn TFA9DNA5srRNA45bpRNA pol
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2transcriptional activation domainDNA80~100AA13 -acidic -helix domainGlnglutamine-tich donainproproline-rich domain
--helix-loop-helix,HLH100~200AA2-1AA2AADNA-CTF/NF-1DNA-AALeucine zipper
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RNA pol2PDNA RNA pol IITATATFIID+TATA RNA pol II TATA+TATAboxPr.-DNApol II pol IIPr.-DNA pol II RNA pol
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1.
12 //cAMP-Pr./ Pr.SerThrSP1DNA
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DNAPr.Vitelinegen,Pr.Pr.DNARNA polPr.DNAPr.2Pr.-PrPr.-Pr.C-myc Pr. Leu max Pr.
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2.Pr. 1Pr. 23. RNA poltrans-acting1looping DNARNA pol 2twisting DNAPr.DNA DNA 3sliding DN4Oozing 14. combinatorial regulation TATATATA box RNA polPr.-DNA open complex
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RNAmRNAtRNAtRNAposttranscription processing5 1.mRNA caping5 GpppmNPtailing AA50~200bp 1mRNA 2CBP 2mRNA 2.rRNA 3.tRNA L
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mRNA 1.splicing)-mRNArRNAtRNAinterveningsequence
1Option exonexon skipping12Option intron 13Mutually exclusiveexon 21mRNA14Internal splice site
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2.1Bax BaxBax/Bcl2 BaxPr.mRNA Bax 6192192AA Bax 5 218 Baz 134562151AA
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2IBr NF- B B NF- BI BrNF B/I Br105KD I BrmRNA178672CPr.I Br1I Br2 21AI Br I BrNF- B
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mRNAmRNA 1.3H-udR20%mRNA RNA1 2.9nm>9nm15nm 20nmgold sphereRNAtRNA5sRNA frogoocyte mRNA 3.RNA20%RNA 2Pr.RNA
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GTP.MET-tRNA40S.MET-tRNAi.GTP 40S.MET-tRNAiGTPmRNA 80s.METtRNA.GTPmRNA 1.Pr. mRNAPr. Pr. mRNA5, Pr.ferritinmRNA530iron-response element 1 1regulatoryiron-binding pro 1Pr.mRNA 2
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2. eIF2Pr. eIF2 eIF2() Rbc 3.5-AuG5 3 mRNA
1Pr.90%mRNA1AUGPr.25AUGAUGCAMP-CAMP-Pr.5-AUG(1)1-AUG
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4.mRNA5 1517~80 21AUG540S121AUGFN5267mRNA mRNA1.1 mRNA3mRNA 2 mRNA10 ,-Pr.mRNA>10h2. 1mRNA 3AUmRNA 2Pr.mRNA3polyADNADNA1h12 mRNA
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3.DNA 1mRNA3mRNA 2Pr.mRNA3Pr.3mRNA 3-Pr.mRNA3mRNA 3mRNA 4 -Pr.mRNA3Pr.mRNA3DNARNAlin-4Lin-14Pr. Lin-42RNA1221340Lin-4RNALin14mRNA3UTRmRNA
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1 Pr. 1.fosmyc 2. 3. 1 2N-1AAMet.serThrAlavalcysGlyproAA12AAAAdestabilizing amino acid AAN-MetMet-tRNAN1AA N1AA/1AA
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Pr.Pr. signal peptide Pr.Pr.
Pr1.1PrN5~10AAPr.H3N-Met---Leu-Leu-Leu-Val-Gly-Ile-Leu-Phe-Trp-Ala---ER21C4AAPr.ER Lys-Asp-Glu-Leu-Coo-
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2.mitochondrion AAAA3~5AA1AAH3N-Met-Leu-Ser-Leu-Arg(+)---3.5AAPro-Pro-Lys-Lys-Lys-Arg-Lys-Val-4. 1FAERSrc Pr.NGlyras Pr.C4cys2src Pr.ras Pr.