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Lecture 9

BIOL442 - Lecture 9 - DNA.pdf

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BIOL 442
Christine Dupont

Set 9 DNACytopathic EffectsCPEChanges in morphological characteristics of cell due to infectionCPE observed in cultured cells for some virusesDiagnosis of viral infections frequently requires the isolation of the virus in cell cultures Cell monolayers are inoculated with a suitable clinical specimen and then observed for cytological changes that indicate virus growthSyncytia are multinucleated giant cells Fused cells multiple nucleiEspecially common with enveloped viruses see example but some naked will do this tooFish Virus LabBe very aware that this is an infectious agent for fishNothing goes down the sink New Section 1 Page 1 Inclusion bodies are another common CPE characteristic of some virusesIntracellular insoluble congregations of usually viral proteins either in cytoplasm or nucleus depending on virusCell has difficulty keeping large amount of viral proteins in properly folded soluble stateReadily stained by certain basic or acidic dyesOther CPEs include rounding up of cells from their normal elongated shapes shrinking or enlargement of cells distortious such as membrane blebbing loss of adherence to culture dish Acheson fig 197 pg 232 reovirus inclusion bodies in infected mouse cell line Immunohistochemical detection with fluorescent antibodiesRabies in a foal brain Neuronal cytoplasmic eosinophilic inclusionsNegri bodiesCanin distemper virus Eosinophilic nuclear inclusion bodiesDNA VirusesAnimalsExcept for Hepadnahas a reverse transcriptase and Poxiciridaewhich is very complex in structure DNA viruses do NOT carry a polymerase with them in the virion TranscriptionEasily done as DNA format is compatible with hostNucleus needed for host DNAdependent RNA polymerase except Poxviridae for transcriptionReplicationDifficulty in relying on DNAdependent DNA polymerase is its need for a primerLinear chromosomes have a problem in replication of the endsIf DNA replication requires many accessory proteins virus is reliant on nucleus and S phaseSimilar to eukaryotes all but the smallest viruses have 2 or 3 origins of replication that support bidirectional synthesisAll viral templates that are transcribed by RNA pol II contain transcriptional control regions that areNew Section 1 Page 2
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