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Lecture8BIO155BB
Course: BIO 155, Fall 2010School: DePaul
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OfDNA BIO155 TheStructureandFunction Dr.JessicaPamment Overview DiscoveryofDNA StructureofDNAandRNA DNAreplication FlowofgeneticinformationfromDNAto RNAtoprotein HistoryTimeline HistoryTimeline 1866GregorMendelshowedtraitsin 1950sDNAidentifiedashereditary material peaswereinheritedindiscretepackages 1953JamesWatsonandFrancisCrick proposedthedoublehelixmodelforDNA DNAandRNAStructure DNAandRNAStructure...
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OfDNA BIO155
TheStructureandFunction Dr.JessicaPamment
Overview
DiscoveryofDNA StructureofDNAandRNA DNAreplication FlowofgeneticinformationfromDNAto
RNAtoprotein
HistoryTimeline HistoryTimeline
1866GregorMendelshowedtraitsin 1950sDNAidentifiedashereditary
material peaswereinheritedindiscretepackages
1953JamesWatsonandFrancisCrick
proposedthedoublehelixmodelforDNA
DNAandRNAStructure DNAandRNAStructure
BothDNAandRNAarenucleicacids Consistofchainsofnucleotides:polynucleotides Nucleotidesarejoinedbycovalentbonds Thephosphategroupiswhatmakesanucleic
acidanacid
The Structure of DNA The
(a) Rosalind Franklin
(b) Franklins X-ray diffraction photograph of DNA
Base Pairing Results in Uniform Diameter Base
Double-ringed base + DRB: too wide Single-ringed base + SRB: too narrow
DRB + SRB: width consistent with X-ray data
Base Pairing by H bonds by
Adenine (A)
Thymine (T)
Guanine (G)
Cytosine (C)
TheDoubleHelix
5 end Hydrogen bond 1 nm 3.4 nm 3 end
3 end 0.34 nm
(a) Key features of DNA structure (b) Partial chemical structure
5 end
(c) Space-filling model
Model for DNA Replication Model
A C T A G T G A T C
(a) Parent molecule
Model for DNA Replication
A C T A G T G A T C A C T A G T G A T C
(a) Parent molecule
(b) Separation of strands
Model for DNA Replication
A C T A G T G A T C A C T A G T G A T C A C T A G T G A T C A C T A G T G A T C
(a) Parent molecule
(b) Separation of strands
(c) Daughter DNA molecules, each consisting of one parental strand and one new strand
DNA Replication Replication
DNAPolymerases
Enzymesthatmakecovalentbonds
betweenthenucleotidesofagrowingDNA strand rateof50nucleotides/sec
Theycatalyzeadditionofnucleotidesata Veryaccurate,withonlyoneinabillion
nucleotidesbeingincorrectlypaired!!!
Damage to DNA by UV light light
ProofreadingandRepairingDNA ProofreadingandRepairingDNA
AccuracyofDNAreplicationisdueto:
1. ProofreadingpicksupmistakesasDNA
isbeingsynthesized
1. DNArepairseveralmechanisms,for
example,nucleotideexcisionrepair (NER)
Nucleotide Excision Repair of DNA damage of
Nuclease
DNA polymerase
DNA ligase
ImportanceofDNArepair ImportanceofDNArepair enzymes
DefectiveDNArepairenzymescanleadto
cancer
XPxerodermapigmentosumarisesfrom
defectiveNERenzymes
Hypersensitivetosunlight Increasedincidenceofskincancer
XerodermaPigmentosum
XPautosomalrecessive disease
Summary Summary
DNAhelixconsistsoftwoantiparallelsugarphosphate
chainsontheoutside,withnitrogenousbasesprojecting intothecentreofthehelix
DNAreplicationissemiconservative ProofreadingandDNArepairmechanismmaintainthe
highfidelityofreplication
BIO155
FromGenetoProtein Dr.JessicaPamment
Overview
Howdoesthegeneticcodegettranslatedto
specificproteins?
Proteinsasthelinkbetweengenotypeand
phenotype
TranscriptionusesDNAtomakemRNA TranslationreadsmRNAtomakeprotein
The Flow of Genetic Information in a Eukaryotic Cell Cell
Transcription and Translation of Codons Codons
GeneticCode
Atripletofbasescorrespondstoeachamino
acid
Thegeneticinstructionsforeachpolypeptideis
writtenasaseriesofthreenucleotidewords
BothDNAstrandscanactastemplates Foreachgene,thesamestrandactsas
templateeverytimeitistranscribed
The Genetic Code The
UniversalityoftheGeneticCode
Thegeneticcodeisnearlyuniversal Allowstransferofgenesfromonespecies
toanother
Exploitedintheareaofbiotechnology Suggestscodeoperatedveryearlyonin
thehistoryoflife,sharedbyallorganisms
(a) Tobacco plant expressing a firefly gene
(b) Pig expressing a jellyfish gene
TranscriptionandTranslation
TranscriptionisthesynthesisofRNAunder
directionofaDNAtemplate
Translationisthesynthesisofapolypeptide
underdirectionofmRNA
Takeplaceinalldomainsoflife
StagesofTranscription
1. Initiation 1. Elongation 1. Termination
RNApolymeraseistheenzymethatcarries outtranscription.
DrawonBoard
The Flow of Genetic Information in a Eukaryotic Cell Cell
Translation:ThePlayers
MessengerRNA(mRNA) TransferRNA(tRNA) Ribosomes
TransferRNA
tRNAisthetranslator tRNArecognizescodonthroughits
anticodon
EachtRNAcarriesaspecificaminoacid
The Structure Structure of tRNA
Ribosomes
BringtogethertRNAandmRNAduring
translation
Madeupoftwosubunits Subunitsjointomakefunctionalribosomein
cytoplasm
The Ribosome The
Translation
Initiation:determineswheretranslation
begins.BringstogethermRNA,thefirst aminoacidwithattachedtRNAandtwo subunits
Elongation Termination
Mutations
The Molecular Basis of Sickle-cell Disease The
Mutations
Achangeinthenucleotidesequenceof
DNA
Usuallyharmfulfortheorganism Butifproteinenhancessuccessof
organism,itllbeselectedbynatural selection
TypesofMutations
1. Substitutionsthereplacementofone
nucleotidepairwithanotherpair ofnucleotidepairsinagene
1. Insertionsanddeletionsadditionsorlosses
Base-Pair Substitution
Wild type
DNA template strand mRNA Protein Stop
T instead of C
A instead of G
Stop Missense
InsertionsandDeletions
Wild type
DNA template strand mRNA Protein Stop
Extra A 3 5 Extra U 5 Stop Frameshift causing immediate nonsense (1 base-pair insertion) 3 5 3
Mutagens
Anagentthatcausesmutations Physicalandchemicalagentsthatinteractwith DNAtocausemutations Mostmutagensarecarcinogenic
TypesofMutagens
i. Chemicalmutagens:intercalating
agents,chemicalsthatalterbasepairing
i. Physicalmutagens:UVlightandXrays
Mutations are the Ultimate Source of of Diversity Life Mutations
Summary
Genesspecifyproteinsviatranscriptionand
translation
TranscriptionistheDNAdirectedsynthesisof
RNAbyRNApolymerase
Transcriptionfollowsthesamebasepairing
rulesasreplication(exception,UinsteadofT)
Summary
Threestagesfortranscriptionandtranslationare
initiation,elongation,andtermination polypeptide onmRNAs
TranslationistheRNAdirectedsynthesisofa tRNAshaveanticodonsthatrecognizecodons
Summary
tRNAsbringinthecorrectaminoacid
specifiedbythecodon
Ribosomescoordinateinitiation,
elongation,andterminationoftranslation
Ribosomes coordinate the 3 stages of translation
mRNA
Ribosome
Polypeptide
Summary
Mutationscanaffectproteinstructureand
function
Conceptofageneisuniversal
BIO155
Viruses Dr.JessicaPamment
CharacteristicsofViruses
Notcellular;noplasmamembrane,cytoplasm,or
ribosomes
Theydontgrow,theyassemble Allcelltypescanbeinfected,butvirusestendto
bespeciesspecific
Theyaresmall Theyareobligateintracellularparasites
T4 bacteriophage infecting an E. coli bacterium
0.5 m
Overview
Whystudyviruses? Thestructureofviruses Viralreproduction Virusesaspathogens
TheImportanceofViruses
Virusesasmodelsystemstostudy
molecularmechanismsofgeneregulation
Virusescauseseriousdiseasesinhumans
andotherorganisms
Virusesastoolsinbiotechnology
StructureofViruses
Virusesareinfectiousparticlesconsistingof
nucleicacidenclosedinaproteincoator membranousenvelope
Sizerange20300nm Components:
1.Viralgenome 2.Proteincoatandenvelopes
ViralGenomes
DNAviruses:doubleorsinglestrandedDNA RNAviruses:doubleorsinglestrandedRNA GenomeisasinglelinearORcircularmolecule Smallestvirushas4genes Largestvirushasseveral100to1000genes
CapsidsandEnvelopes
Acapsidisaproteinshellenclosingthe
genome
Capsidsareshapeddifferentlydepending
onvirus
Membranousenvelopesurroundingsome
capsidsarederivedfromhostcell
Viral Structure
RNA DNA RNA DNA
18 250 nm
7090 nm (diameter)
80200 nm (diameter)
80 225 nm
20 nm
50 nm
50 nm
50 nm
(a) Tobacco mosaic virus
(b) Adenoviruses
(c) Influenza viruses
(d) Bacteriophage T4
ViralReproductiveCycles
Virusescanonlyreproduceinhostcells Eachtypeofviruscanonlyinfectitshostrange Virusesidentifyhostcellsbylockandkey Entryintohostcellcanbebyendocytosisorby
fusionwithplasmamembrane
Somevirusesinjectnucleicacidintocell
Bacteriophages infecting an E. coli cell E.
ReproductiveCyclesofPhages
2alternativemechanisms: andlysisofhostcell theirhostcell
1. Lyticcycleusedbyvirulentphages.Resultsindeath 1. Lysogeniccycleusedbyphagesthatcoexistwith
Temperatephagescanusebothmodesof reproduction
The Lytic Cycle The
Alternative Phage Reproductive Cycles Alternative
ViralDiseasesinPlants
Over2000viraldiseaseinplantsknown Causelossofcrops MostplantvirusesareRNAviruses
Tobacco Mosaic Virus Tobacco
Viral Infection in Plants
AnimalViruses
Classifiedonbasisofgenome MostRNAviruseshaveanenvelope
Herpes Simplex 1 Herpes
HIV
HumanImmunodeficiencyVirus AretrovirusthatcausesAIDS(Aquired
ImmunoDeficiencySyndrome)
Envelopedvirus ContainstwoidenticalssRNAmolecules+tworeverse
transcriptases
InformationflowsfromRNAtoDNA Provirusneverleavesthehostcell
VideosonHIVLifeCycle
http://www.metacafe.com/watch/334765/hiv_lif http://www.metacafe.com/watch/yt
e7ydwpOah3I/hiv_life_cycle/
AIDS
AcquiredImmunodeficiencySyndrome Duetoweakenedimmunesystem,secondary
infectionsusuallykillAIDSpatients
TwomaintypesofantiHIVdrugs: 1.AZTinhibitsreversetranscriptase 2.Proteaseinhibitingdrugs ProblemisHIVsrapidrateofmutation!
EmergingViruses
Newviruses,suchasHIV,Ebola,SARS Processesthatcontributetoemergenceof newviruses:
1. Mutationofexistingviruses 1. Disseminationofviraldiseases 1. Spreadofexistingvirusesbetweenanimal
species
SARS
SevereAcuteRespiratorySyndrome FirstreportedinChinain2003 Within3monthsover8,000peopleinfected CoronavirusresponsibleforSARShadnever
beforecauseddiseaseinhumans
Mutationslikelycauseforitsemergence
SARS Virus SARS
RNAvirus
InfluenzaViruses
3types:A,BandC BandCinfectonlyhumans,nevercausedanepidemic TypeAfoundinbirdsandsomemammals InfluenzaAcausesnosymptomsinwildbirds InfluenzaAcausessymptomsindomesticbirdsand
rarelyinhumans
InfluenzaAisthecauseof3majorepidemics
InfluenzaAVirus
Classificationbasedonviralsurfaceproteins,HandN 16typesofHand9typesofN H5N1straincrossedspeciesbarrierfrombirdsto
humanscausingavianflu
Highmortalityrate Latestflupandemicistheswineflu(H1N1)firstdetected
inUSinApril2009.Containspig,birdandhumanflu genes
RearrangementofViralGenomes
Therearehuman,horse,pig,chicken,andduckflu
viruses
Ordinarilydontcrossinfect Ifhumanwiththefluisinfectedwithflufromdifferent
species,RNAcanrearrange virulentdiseases
Crossspeciesrearrangementisthecauseofmost Spanishfluwasevenworseasthereappearstohave
beennorearrangementwithhumanvirus,makingit completelyunrecognizabletoIS
Influenza in animals
(a) The 1918 flu pandemic
0.5 m
(b) Influenza A H5N1 virus
(c) Vaccinating ducks
Summary
AvirusconsistsofRNAorDNAsurroundedbyaprotein
coat
Virusesareacellularparasites Virusesreproduceonlyinhostcells Lyticandlysogeniccyclesofbacteriophages ReproductivecyclesofHIV,ananimalvirus
GeneDefinition
Unitofinheritance Regionofspecificnucleotidesequence AgeneisaregionofDNAthatcanbe
expressedtoproduceafinalproducethat iseitherapolypeptideoranRNAmolecule
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