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VIRUSSTRUCTURE
Virionvsvirus
Virionistheinfectious
particle
composedofnucleic
acid,proteincapsid,
+/envelope
maybeextracellularor
intracellular
Virusisanystageof
infection
HowdoweknowthatNAisgenetic
material?
HersheyChase FraenkelConrat
Experiment
TRANSFECTION
EXPTS
TRANSFECTIONFAILSFOR
SOMEVIRUSES
WHY?
Capsid
Functions
ProtectionofNA
Attachmentfornaked
viruses
Enzyme
HelicalvsIcosahedral
SymmetryWhydomost
viruseslookalike?
Tobacco mosaic virus is a
ssRNA virus composed of
6000 nucleotides. The capsid
is made of 2100 copies of a
single protein subunit that
contain 158 amino acids.
Calculate the percentage of
the genome that is used for
structure.
Howdohelicalvirusesdiffer?
Helicaloneaxisofsymmetry
downcenter
Multiplestructuralunits
Icosahedralsymmetry
20identicalequilateraltriangles
Structuralunitsonfacesto
givemorphologicalcapsomers
Pentons(5foldaxisof
symmetry)
Hexons
3foldthroughface
2foldthroughedge
Howdospherical
virusesdiffer?
Envelope
Attachment
Entry
Assemblymatrix
proteins
Release
Proteinsareviral
Lipidsarehost
Rareinplantsorbacteria
why?
If the membrane envelope is
destroyed, the virus becomes
noninfectious. Why?
Herpesviruscomplexity
Tegumentproteins12/84viral
proteinsinHSV
Potentialrole?
VirionmRNA
DNAasevirionnucleic
acids
RTPCR
probegenomearray
Potentialrole?
GenomeDNA
Genome or RNA
RNAGenomes
sense (positive-sense,
negative-sense,
ambisense)
presence or absence of 5'-
terminal cap or 5'-
covalently-linked protein
presence or absence of 3'-
terminal poly (A) tract
Retroviruses - replication
strategy
Someviruseshavehighdegreeofsecondary
structure
Poliovirus5internal
ribosomeentrysite(IRES)
Guestetal.2004.J.Virol.78:11097.
SARS/coronaviruseshaveconserved3region
SARSs2minred
agreen=530loopof16SRNA
Similarbindingproperties:
bblue=S12
magenta=IF1
Possiblerolefors2m
Hijacksproteinsynthesisfrom
cell(bindingcellfactors)
Neededtobindtosimilarviral
proteinfortranscription
Potentialdrugtargetinredtunnel
Robertsonetal.2005.PLOsBiology:3.
DNAVirusesmaybelargegenomes
PolyDNAvirus(PDV)contain
manyDNAsegments
Mimiviruslargerthansmall
bacteria
Hostinducedmodification
Viralpropertythatvaries
dependingonthehost
PhageDNAhydroxymethyl
cytosine(HMC)replacesC
Viralenzymes:CtoHMC
ViralDNApolymerase:adds
HMCnotC
WhatisadvantageofHMC?
GlucoseisattachedtoHMC
Hostenzymeneededto
prepareglucose
Protectsagainsthostnuclease
Whatwouldhappenif
viruswithoutglucose
entershostwithRE?
Whatwouldhappenif
viruswithglucose
entershostw/o
enzymetocreate
Hostenzymemakes UDPglucose?
Proteins
structuralproteins
nonstructuralvirion
proteins
transcriptase,
protease
integrase
Howtoidentifyvirionproteins
PurifyKSHVvirions
RunonSDSPAGE
Excisebands,digestget
sequenceandcompareto
database
Chemicalsynthesisofpoliovirus:Whatare
theimplications?
SmallgenomepositivestrandRNAsequenceknown
SynthesizedsmallDNAsegments(~69nucleotides)with
overlappingcomplementarysegments
AddedaT7phagepromotortoDNA
UsedDNAtomakegenomeRNAinHeLacelllysatewith
T7polymerase
Results:Howdoyoushowsuccess?
QuickTime and a
TIFF (Uncompressed) decompressor
are needed to see this picture.
InternationalCongressonTaxonomyofViruses
http://www.ncbi.nlm.nih.gov/ICTV/
Morphology Genomecharacteristics
virionsize Replicationstrategy
envelopedornaked Antigenic Properties
nucleocapsid
capsidsymmetryand
structure
Baltimoreclassification
WHYTRANSFECTIONFAILS
ONESTEPGROWTHCURVE
1939EllisandDelbruck:
Infectionwithahighmultiplicity
ofinfection(MOI):ratioofvirus
tohostcell
Simultaneousinfection
Singlereplicationcycle
Sampleattimeintervalsbyplaque
countforplaqueformingunits
(PFU),
Identificationoflatentphase
Determinationofburstsize/viral
yield
MeasuringIntracellularEvents
Sampleattimeintervals
afterlysingcells(1952
Doermann)
Chloroform
Lysisfromwithout
Identificationofeclipse Maturationphase
andmaturationphases