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7/29/2019 10-Reg Euc.pdf
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7/29/2019 10-Reg Euc.pdf
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Facts:
- Bacteria(4000genes)
90%genesareinvolvedwithgrowthandprolifera=on10%genesdifferen=a=on
havegenesorganizedinoperons
promotersaresmallandveryspecific
- Yeasts(4500genes)
90%ofgenesareinvolvedwithgrowthandprolifera=on
10%ofgenesdifferen=a=onhavegenesdispersed
promotersaremediumsizeandveryspecific
- Mul=cellular(25000genes)
20%genesinvolvedwithgrowthandprolifera=on
40%genesinvolveswithdevelopment
40%genesinvolvedwithspecializedfunc=ons
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+
Howisamul=cellularorganismconstructed?
Highlyspecializedcells
zygote
To=potentcell
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Docellsloosegenesastheyareundergoing
developmentanddifferen=a=on?
CloningsaysNO!!
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Expressionofdifferentgenesindifferentcellstypes
DNAmicro-arrays
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Differentproteinsareproducedindifferent
=ssuesatdifferent=mes
2D-SDSproteingelelectrophoresis
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Regula=onofgeneexpressioncanoccuratmanydifferentstages
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Threekeycomponentsintheregula=onofgeneexpression:
1. Signals: hormones(estrogen,testosterone,progesterone) Growthfactors(NGF,CSF,Interferon,…)
environment(drugs,toxicsubstances,metals)
2.Levelofresponse
transcrip=on
mRNAprocessing
mRNAtransportandlocaliza=on
mRNAtransla=on
mRNAstability
Proteinac=vity
3.Mechanisms
Transac=va=ngproteins
Methyla=on
X-inac=va=on
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Transcrip=onisini=atedbyproteinsrecognizingthePromoterofagene
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Transcrip=onfactorsbindspecificregulatorycis-ac=ngsequencesand
promotetranscrip=on
mRNA+1
TATA
-5000pb
RNApolyII
mRNA
TATA
-5000pb
“OFF”
“low”
mRNA
TATA
-5000pb“ON”
EnhancerBasic
promoter Transcrip=onfactors
+1
+1
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Transcrip=onfactorsrecogniseshortDNAsequences
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FamiliesofDNA-bindingtranscrip=onfactors
-DNAbinding
proteins
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Differenttypesofregula=on
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Organiza=onofaeukaryo=cpromoter
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Organiza=onofaeukaryo=cgeneregulatoryregion
GeneTATA-5000pb
Enhancer5’ Enhancer3’
BasicPromoter
Basicpromoter
hasfixed:
-posi=on
-distance
-orienta=on
Enhancers
havevariable:
-distance
-posi=on
-orienta=on
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Ac=vityofenhancersatadistance
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Ac=vityofenhancersatadistance
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Sequenceofeventsduringac=va=onoftranscrip=on
Histoneacetyla=onàgenetranscrip=on
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Assemblyoftranscrip=onalcomplexesatthepromoter
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Possiblepost-transla=onalmodifica=onsoftranscrip=onfactors
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Differen=alexpressionoffetalandmaternalglobingenes
γGγA
ε
β
δ
LowOxigenpressure,highaffinity
HighOxigenpressue,loweraffinity
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Glucoccor=coidsdiffusetothenucleusandcouldworkatmanydifferentpromoters
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Alterna=vesplicing:howtogetmoreproteinsoutof
yourgenome
30,000humangenesprobablygivemorethan100,000differentproteins
21 3 4calcitonin 5CGRP 6
5’ 3’
21 3 4calcitonin
21 3 4calcitonin 5CGRP 6
21 3 4calcitonin
21 3 5CGRP 6
AAAA…
AAAA…
DNA
Nuclear
Transcripts
mRNAs
Proteins
Thyroid
Neurons
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Alterna=vepromotersandsplicinga-amylase
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Regula=ngstabilityofmRNA
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Promoterdele=onstudies
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Promoterdele=on
studieswithGFPin
Wholeanimals
Conec=ve=ssuegrowth
factor
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CellularStress
Response
Manygenesupregulated
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ControlofGALregula=oninyeast
- Ac=vatorproteinGAL4
- InhibitoryproteinGAL80
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Eukaryo=cpromotersareverycomplexandreceivemul=plesignals
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0
Complexityofeukaryo=cpromoters
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Nuclearmatrixassocia=onofpromoters
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GenomawidemappingSARs/MARs
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Natureofinsulators
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Theeffectofcytosinemethyla=onupongeneexpression
HeritablepaernsofexpressionmaintainedthoughDNAreplica=on
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Theeffectofcytosinemethyla=onupongeneexpression
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Housekeepinggenes
CpGrepe==veblocksusuallymarkgenescons=tu=velyexpressed
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DrosophilaDevelopmentalGene=cs
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Combinatorialgenecontrolduringdevelopment
b l l d d l
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Combinatorialgenecontrolduringdevelopment
Th i = l d
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Theepigene=clandscape
ConradWaddington(1905-1975)
Th i = l d
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Theepigene=clandscape
ConradWaddington(1905-1975)
Th i = l d
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Theepigene=clandscape
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Epigene=cgeneregula=on