版权说明:本文档由用户提供并上传,收益归属内容提供方,若内容存在侵权,请进行举报或认领
文档简介
GeneExpressionOverview
SalwaHassanTeama
1
GeneExpressionTheGeneStructureTranscriptionGeneticCodeandProteinSynthesisRegulationofGeneExpressionProkaryotesVsEukaryotesGeneExpressionAnalysis2ContentsTargetaudiencePhysicianassistant;Postgraduateinclinicalspecialties;Medicalstudents;Medicaltechnologist;Beginners;andForeverylaboratoryworkerandeveryonepassionforlearning3GeneExpressionTheprocessbywhichagene'sinformationisconvertedintothestructuresandfunctionsofacellbyaprocessofproducingabiologicallyfunctionalmoleculeofeitherproteinorRNA(geneproduct)ismade.Geneexpression
isassumedtobecontrolledatvariouspointsinthesequence
leadingtoproteinsynthesis.
4GeneStructure
Eukaryoticgenestructure:Mosteukaryoticgenesincontrasttotypicalbacterialgenes,thecodingsequences(exons)areinterruptedbynoncodingDNA(introns).Thegenemusthave(Exon;startsignals;stopsignals;regulatorycontrolelements).Theaveragegene7-10exonsspreadover10-16kbofDNA.56ProteinSynthesis:FourstagesTranscriptionRNAprocessingTranslationPost-translationprocessing78GeneExpressionTranscriptionSynthesisofanRNAthatiscomplementarytooneofthestrandsofDNA.TranslationRibosomesreadamessengerRNAandmakeproteinaccordingtoitsinstruction.9ProteinSynthesis10Transcription
11TranscriptionEnzymesRNApolymerase:Theenzymethatcontrolstranscriptionandischaracterizedby:SearchDNAforinitiationsite,ItunwindsashortstretchofdoublehelicalDNAtoproduceasingle-strandedDNAtemplate,Itselectsthecorrectribonucleotideandcatalyzestheformationofaphosphodiesterbond,Itdetectsterminationsignalswheretranscriptends.12EukaryoticRNApolymeraseshavedifferentrolesintranscription
PolymeraseInucleolusMakesalargeprecursortothemajorrRNA(5.8S,18Sand28SrRNAinvertebratesPolymeraseII
nucleoplasmSynthesizeshnRNAs,whichareprecursorstomRNAs.ItalsomakemostsmallnuclearRNAs(snRNAsPolymeraseIIINucleoplasmMakestheprecursorto5SrRNA,thetRNAsandseveralothersmallcellularandviralRNAs.EukaryoticPromoterConservedeukaryoticpromoterelementsConsensussequenceCAATboxGGCCAATCTTATAboxTATAAGCboxGGGCGGCAPsiteTAC14
EukaryoticPromoterliesupstreamofthegene.Thereareseveraldifferenttypesofpromoterfoundinhumangenome,withdifferentstructureanddifferentregulatorypropertiesclass/I/II/III.TranscriptionFactorsTranscriptionfactorsareproteinsthatbindto
DNAnearthestartoftranscriptionofagene.TranscriptionfactorseitherinhibitorassistRNApolymeraseininitiationandmaintenanceoftranscription.15TranscriptionFactors16EnhancersEnhancersarestretchesofbaseswithinDNA,about50to150basepairsinlength;theactivitiesofmanypromotersaregreatlyincreasedbyenhancers
whichcanexerttheirstimulatoryactionsoverdistancesofseveralthousandsbasepairs.17PreinitiationComplexThegeneraltranscriptionfactorscombinewithRNApolymerase
toformapreinitiationcomplexthatiscompetenttoinitiatetranscriptionassoonasnucleotidesareavailable.
Theassemblyofthepreinitiationcomplexoneachkindofeukaryoticpromoter(classIIpromotersrecognizedbyRNApolymeraseII)
beginswiththebindingofanassemblyfactortothepromoter.1819Source:/health/What-is-Gene-Expression.aspx20Initiation
ThepolymerasebindingcausestheunwindingoftheDNAdoublehelixwhichexposeatleast12basesonthetemplate.
ThisisfollowedbyinitiationofRNAsynthesisatthisstartingpoint.21Initiation
TheRNApolymerase
startsbuildingtheRNAchain;itassemblesribonucleotidestriphosphates:ATP;GTP;CTPandUTPintoastrandofRNA.Afterthefirstnucleotideisinplace,thepolymerasejoinsasecondnucleotidetothefirst,formingtheinitialphosphodiesterbondintheRNAchain.22Elongation
RNApolymerase
directsthesequentialbindingofriboncleotidestothegrowingRNAchaininthe5'-
3'direction.EachribonucleotideisinsertedintothegrowingRNAstrandfollowingtherulesofbasepairing.ThisprocessisrepeatedutillthedesiredRNAlengthissynthesized……..23Termination
Terminators
attheendofgenes;signaltermination.TheseworkinconjunctionwithRNApolymerasetoloosentheassociationbetweenRNAproductandDNAtemplate.TheresultisthattheRNAdissociatefromRNApolymeraseandDNAandsostoptranscription.TheproductisimmatureRNAorpremRNA(Primarytranscript).24TheprimaryproductofRNAtranscription;thehnRNAscontainbothintronicandexonicsequences.ThesehnRNAsareprocessedinthenucleustogivematuremRNAsthataretransportedtothecytoplasmwheretoparticipateinproteinsynthesis.2526RNAProcessing(Pre-mRNA→mRNA)CappingSplicing
AdditionofpolyAtail27RNAProcessingCappingThecapstructureisaddedtothe5'ofthenewlytranscribedmRNAprecursorinthenucleuspriortoprocessingandsubsequenttransportofthemRNAmoleculetothecytoplasm.Splicing:
StepbystepremovalofpremRNAandjoiningofremainingexons;ittakesplaceonaspecialstructurecalledspliceosomes.28RNAProcessingAdditionofpolyAtail:Synthesisofthepoly(A)tailinvolvescleavageofits3'endandthentheadditionofabout40-200adenineresiduestoformapoly(A)tail.2930AlternativeSplicingAlternativesplicing:
isaverycommonphenomenoninhighereukaryotes.Itisawaytogetmorethanoneproteinproductoutofthesamegeneandawaytocontrolgeneexpressionincells.3132Source:/Class/MLACourse/Modules/MolBioReview/alternative_splicing.htmlTheGeneticCodeThesequenceofcodonsinthemRNAdefinestheprimarystructureofthefinalprotein.ThreenucleotidesinmRNA(acodon)specifyoneaminoacidinaprotein.33TheGeneticCode34TheGeneticCodeThetripletsequenceofmRNAthatspecifycertainaminoacid.64differentcombinationofbases;61ofthemcodefor20aminoacids(AA);thelastthreecodon(UAG,UGA,UAA)donnotcodeforaminoacids;theyareterminationcodons.Degenerate
Morethanontripletcodonspecifythesameaminoacid.35TheGeneticCodeUnambiguousEachcodonspecifiesaparticularaminoacid,thecodonACGcodesfortheaminoacidthreonine,andonlythreonine.NonoverlappingThismeansthatsuccessivetripletsarereadinorder.Eachnucleotideispartofonlyonetripletcodon.36DNACodon
RNACodon
37
Translation
38TranslationTranslation
istheprocessbywhichribosomesreadthegeneticmessageinthemRNAandproduceaproteinproductaccordingtothemessage'sinstruction.39RequirementforTranslation
RibosomestRNAmRNAAminoacidsInitiationfactorsElongationfactorsTerminationfactorsAminoacyltRNAsynthetaseenzymes:Energysource:40RibosomesEukaryoticribosomesarelarger.Theyconsistoftwosubunits,whichcometogethertoforman80Sparticle;60Ssubunitholds(threerRNAs5S,5.8S,28Sandabout40proteins).40Ssubunitcontains(an18SrRNAandabout30proteins).41ThelargeribosomalsubunitcontainsthreetRNAbindingsites,designatedA,P,andE.
TheAsite
bindsanaminoacyl-tRNA(atRNAboundtoanaminoacid);Psite
bindsapeptidyl-tRNA(atRNAboundtothepeptidebeingsynthesized).The
Esite
bindsafreetRNAbeforeitexitstheribosome.42PreparatoryStepsforProteinSynthesis
First,aminoacyltRNAsynthetasejoinsaminoacidtotheirspecifictRNA.Second,ribosomesmustdissociateintosubunitsattheendofeachroundoftranslation.43Theproteinsynthesisoccurin3phases
Accurateandefficientinitiation
occurs;theribosomesbindstothemRNA,andthefirstaminoacidattachedtoitstRNA.Chain
elongation,
theribosomesaddsoneaminoacidatatimetothegrowingpolypeptidechain.Accurateandefficienttermination,
theribosomesreleasesthemRNAandthepolypeptide.44Initiation
Theinitiationphaseofproteinsynthesisrequiresover10eukaryoticInitiationFactors(eIFs):
Factorsareneededtorecognizethecapatthe5'endofanmRNAandbindingtothe40sribosomalsubunit.BindingtheinitiatorMet-tRNAiMet(methionyl-tRNA)tothe40Ssmallsubunitoftheribosome.45InitiationScanningtofindthestartcodonbybindingtothe5'capofthemRNAandscanningdownstreamuntiltheyfindthefirstAUG(initiationcodon).ThestartcodonmustbelocatedandpositionedcorrectlyinthePsiteoftheribosome,andtheinitiatortRNAmustbepositionedcorrectlyinthesamesite.OncethemRNAandinitiatortRNAarecorrectlybound,the60Slargesubunitbindstoform80sinitiationcomplexwithareleaseoftheeIFfactors.46Elongation
Transferofproperaminoacyl-tRNAfromcytoplasmtoA-siteofribosome;Peptidebondformation;PeptidyltransferaseformsapeptidebondbetweentheaminoacidinthePsite,andthenewlyarrivedaminoacyltRNAintheAsite.Thislengthensthepeptidebyoneaminoacids.47ElongationTranslocation;translocationofthenewpeptidylt-RNAwithitsmRNAcodonintheAsiteintothefreePsiteoccurs.NowtheAsiteisfreeforanothercycleofaminoacylt-RNAcodonrecognitionandelongation.EachtranslocationeventmovesmRNA,onecodonlengththroughtheribosomes.48Termination
Translationterminationrequiresspecificproteinfactorsidentifiedasreleasingfactors,RFsinE.coliandeRFsineukaryotes.Thesignalsforterminationarethesameinbothprokaryotesandeukaryotes.ThesesignalsareterminationcodonspresentinthemRNA.Thereare3terminationcodons,UAG,UAAandUGA.49Termination
Aftermultiplecyclesofelongationandpolymerizationofspecificaminoacidsintoproteinmolecules,anonsensecodon=terminationcodonofmRNAappearsintheAsite.Theisrecognizedasaterminalsignalbyeukaryoticreleasingfactors(eRF)whichcausethereleaseofthenewlysynthesizedproteinfromtheribosomalcomplex.50PolysomesMostmRNAaretranslatedbymorethanoneribosomeatatime;theresult,astructureinwhichmanyribosomestranslateamRNAintandem,iscalledapolysomes.51ControlofGeneExpressionTranscriptional
Posttranscriptional
Translational
Posttranslational
52ControlofGeneExpression53Controlofgeneexpressiondependsvariousfactorsincluding:
Chromosomalactivationordeactivation.Controlofinitiationoftranscription.ProcessingofRNA(e.g.splicing).ControlofRNAtransport.ControlofmRNAdegradation.Controlofinitiationoftranslation(onlyineukaryotes).Post-translationalmodifications.5455GeneExpressionAnalysisPolymeraseChainReactionQuantitativePCRMicroarray…….……56EukaryoticGeneExpression
Essentiallyallhumans'genescontainintrons.Anotableexceptionisthehistonegeneswhichareintronless.Eukaryotegenesarenotgroupedinoperons.Eacheukaryotegeneistranscribedseparately,withseparatetranscriptionalcontrolsoneachgene.EukaryoticmRNAismodifiedthroughRNAsplicing.EukaryoticmRNAisgenerallymonogenic(monocistronic);codeforonlyonepolypeptide.57EukaryoticGeneExpression
EukaryoticmRNAcontainnoShine-Dalgarnosequencetoshowtheribosomeswheretostarttranslating.Instead,mosteukaryoticmRNAhavecapsattheir5`endwhichdirectsinitiationfactorstobindandbeginsearchingforaninitiationcodon.EukaryoteshaveaseparateRNApolymeraseforeachtypeofRNA.Ineukaryotes,polysomesarefoundinthecytoplasm.EukaryoticproteinsynthesisinitiationbeginswithmethioninenotNformyl-methionine.58Prokaryoticvs.EukaryoticBacterialgeneticsaredifferent.Prokaryotegenesaregroupedinoperons.ProkaryoteshaveonetypeofRNApolymeraseforalltypesofRNA,mRNAisnotmodifiedTheexistenceofintronsinprokaryotesisextremelyrare.59Prokaryoticvs.EukaryoticToinitiatetranscriptioninbacteria,sigmafactorsbindtoRNApolymerases.RNApolymerases/sigmafactorscomplexcanthenbindtopromoterabout40deoxyribonucleotidebasespriortothecodingregionofthegene.Inprokaryotes,thenewlysynthesizedmRNAispolycistronic(polygenic)(codeformorethanonepolypeptidechain).Inprokaryotes,transcriptionofageneandtranslationoftheresultingmRNAoccursimultaneously.Somanypolysomesarefoundassociatedwithanactivegene.60GlossaryAllelesareformsofthesamegenewithsmalldifferencesintheirsequenceofDNAbases.Alternativesplicing:isaverycommonphenomenoninhighereukaryotes.Itisawaytogetmorethanoneproteinproductoutofthesamegeneandawaytocontrolgeneexpressionincells.Exon:asegmentofagenethatisrepresentedinthematureRNAproduct.IndividualexonsmaycontaincodingDNAand/ornoncodingDNA(untranslatedsequences).Bioinformatics
I
istheapplicationof
computerscience
and
informationtechnologytothefieldof
biologyand
medicine
Introns(interveningsequence)(AnoncodingDNAsequence):InterveningstretchesofDNAthatseparateexons.Primarytranscript:Theinitialproductionofgenetranscriptioninthenucleus;anRNAcontainingcopiesofallexonsandintrons.RNAgeneornon-codingRNAgene:RNAmoleculethatisnottranslatedintoaprotein.NoncodingRNAgenesproducetranscriptsthatexerttheirfunctionwithouteverproducingproteins.Non-codingRNAgenesincludetransferRNA(tRNA)andribosomalRNA(rRNA),smallRNAssuchassnoRNAs,microRNAs,siRNAsandpiRNAsandlastlylongncRNAs.Enhancersandsilencers:areDNAelementsthatstimulateordepressthetranscriptionofassociatedgenes;theyrelyontissuespecificbindingproteinsfortheiractivities;sometimesaDNAelementscanacteitherasanenhancerorsilencerdependingonwhatisboundtoit.Activators:Additionalgene-specifictranscriptionfactorsthatcanbindtoenhancerandhelpintranscriptionactivation.Openreadingframe(ORF):Areadingframethatisuninterruptedbytranslationstopcodon(readingframethatcontainsastartcodonandthesubsequenttranslatedregion,butnostopcodon).Directionality:inmolecularbiology,referstotheend-to-endchemicalorientationofasinglestrandofnucleicacid.Thechemicalconventionofnamingcarbonatomsinthenucleotidesugar-ringnumericallygivesrisetoa5'endanda3'end("fiveprimeend"and"threeprimeend").Therelativepositionsofstructuresalongastrandofnucleicacid,includinggenes,transcriptionfactors,andpolymerasesareusuallynotedasbeingeitherupstream(towardsthe5'end)ordownstream(towardsthe3'end).ReverseTranscription:Someviruses(suchasHIV,thecauseofAIDS),havetheabilitytotranscribeRNAintoDNA.Pseudogenes.DNAsequencesthatcloselyresembleknowngenesbutarenonfunctional.More:/books/NBK7584/61ReferencesandFurtherReadingAliKhalifa.Appliedmolecularbiology;eds:(FathiTashandSannaEissa).109pages.Egypt.UniversityBookCenter.2002.AvailableinpapercopyfromthepublisherDanielH.Farkas.DNASimplified:TheHitchhiker'sGuidetoDNA.110pages.Washington,DC:AACCPress,1996,ISBN0-915274-84-1.AvailableinpapercopyfromthepublisherDanielP.Stites,AbbaT.Terr.BasicHumanImmunology:336Pages.Appleton&Lange(November1990).ISBN.0838505430.AvailableinpapercopyfromthepublisherInnis,DavidH.Gelfand,John
温馨提示
- 1. 本站所有资源如无特殊说明,都需要本地电脑安装OFFICE2007和PDF阅读器。图纸软件为CAD,CAXA,PROE,UG,SolidWorks等.压缩文件请下载最新的WinRAR软件解压。
- 2. 本站的文档不包含任何第三方提供的附件图纸等,如果需要附件,请联系上传者。文件的所有权益归上传用户所有。
- 3. 本站RAR压缩包中若带图纸,网页内容里面会有图纸预览,若没有图纸预览就没有图纸。
- 4. 未经权益所有人同意不得将文件中的内容挪作商业或盈利用途。
- 5. 人人文库网仅提供信息存储空间,仅对用户上传内容的表现方式做保护处理,对用户上传分享的文档内容本身不做任何修改或编辑,并不能对任何下载内容负责。
- 6. 下载文件中如有侵权或不适当内容,请与我们联系,我们立即纠正。
- 7. 本站不保证下载资源的准确性、安全性和完整性, 同时也不承担用户因使用这些下载资源对自己和他人造成任何形式的伤害或损失。
最新文档
- 光电测速仪课程设计
- 光照日晷课程设计
- 光控灯设计课程设计
- 光学工程本科课程设计
- 假花施工方案
- 人工操场施工方案
- 临沂冲孔围挡施工方案
- 中坚外墙真石漆施工方案
- 个人客户营销课程设计
- 上海钢带管施工方案
- 碳中和技术概论全套教学课件
- 《义务教育道德与法治课程标准(2022年版)》
- 2024年中国融通集团招聘笔试参考题库含答案解析
- 部编本语文四年级上册第三单元教材解读-PPT
- 精品课程《人文地理学》完整版
- GA 667-2020防爆炸透明材料
- 教师课堂管理技巧课件
- 【教师必备】部编版五年级语文上册第二单元【集体备课】
- 公司员工待岗、离岗、解聘(解除劳动合同)
- 广州市公共建筑节能设计要求审查表
- 劳动竞赛分包单位责任状wwww
评论
0/150
提交评论