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Virology Intro, RNA Viruses -more prone to mutations -less proofreading activity Sense (-)RNA Viruses, Segmented Genome, Rep in Nucleus -eg. Influenza virus -segmented genome->some segments spliced -replication in nucleus -steals mRNA cap from cellular mRNA -mutations->antigenic drift->flu pandemics, DNA Viruses Types/Location Cytoplasmic DNA Viruses-Poxviruses -carries DNA-dep RNA pol -encodes all replication enzs -initial partial uncoating->limited gene exp ->triggers further uncoating, DNA release -also has DNA rep, RNA transcription enzs ->DNA rep, structural proteins -virus assembled in cytoplasm -builds own membrane ->lysis (not budding), Common Steps 1. Attachment 2. Penetration 3. Uncoating 4. Macromolecular Synth 5. Assembly 6. Release - budding/lysis Steps Assembly/Release Linked processes (influenza) -viral glycoproteins into cell memb ->M proteins associate w/ GPs, genome -modified memb region buds Independent processes (Adenovirus, Herpes) -HV capsid assembly like AV, but envelopes, DNA Viruses Types/Location ssDNA Viruses - Parvovirus -Linear genome, hairpin loops ->converted to dsDNA via cell's S phase -use cell's RNA pol II for transcription -released by cell lysis eg AAV-can complementate w/ Adenovirus, Common Steps 1. Attachment 2. Penetration 3. Uncoating 4. Macromolecular Synth 5. Assembly 6. Release - budding/lysis Steps Attachment-may require ɭ VAP/rec -viral attachment proteins -on capsid or envelope -receptors-NL cell surface proteins -VAP-receptor interaction determines tissue tropism, host range, Viruses Diagnostic Virology Handling -collect from right source -protein, antiBx transfer medium -Refrigerate at 4C -send to lab, Viruses Structure Genome DNA-ss or ds -linear or circular RNA-(+) or (-) sense -ss or ds -continuous or segmented Proteins for protection of other functions - e.g. replication, Viruses Diagnostic Virology Methods: Non-culture-in clinical samples Cytology-inclusion bodies EM IF, EIA for viral Ags Detections of Nucleic acid -via in situ hybridization -or PCR Culture Cytopathic Effects (->plaque) -via Cell culture + Virus -IF to see viral Ags Viral Serology-via pt's ABs -4-fold increase in AB titer -only used if others fail -EBV, Hep, HIV-1, HTLV, arboviruses, RNA Viruses -more prone to mutations -less proofreading activity Sense (-)RNA Viruses w/ Segmented Genome -eg Bunyavirus, Hantavirus -ɯ segments-L, M, S -replicate in cytoplasm -mRNA transcribed w/ virion's RNA-dep RNA pol -can have "ambisense" genome segments, DNA Viruses Types/Location Nuclear dsDNA Viruses -use cell's RNA pol II, xscript proteins ->makes mRNA Immediate early phase - herpesvirus -viral proteins req'd to trigger viral gene exp, rep Early phase-enzs for genome replication -uses virus/cell's enzs for genome rep -may trick cell into entering cell cycle Late phase-enzs for structural proteins -after genome copied -capsid assembly in nucleus eg Papilloma, Polyoma, Adeno - released by lysis -HPV-point mutations change E6, E7 genes -diff affinities to Rb, p53-high vs low risk Molecular Piracy of cellular genes - eg KSHV