MCD – Microbiology 1 – Virus Properties Anil Chopra 1. Describe the nature of viruses: their small size, mode of replication and diversity. Viruses Viruses are small in size, 20-450 nm They are obligate (cannot survive outside host) intracellular parasites. Composition: nucleic acid (DNA or RNA, can be single stranded or double stranded, linear or circular) and protein, (sometimes lipid and carbohydrate too) They have a unique mode of replication. Diversity: all species are infected by viruses, may cause great plagues or be asymptomatic Classification according to: o Type of disease o Mode of transmission o Structure o Immunological relatedness o Nucleic acid sequence o Mode of replication. Viruses are measured in various ways including: o Observing disease in host o Plaque assay (infectivity) – A selection of susceptible host cells is infected with the virus. An area of killed cells large enough to be seen = plaque. o Electron microscopy o Polymerase chain reaction o Immunological evidence of infection 2. Examples of virus replication HIV, polio, influenza, herpes simplex virus Virus replication occurs in a series of steps. This includes: Binding to host cell: often very specific interaction between virus surface proteins and cell receptors HIV gp120 : CD4 Epstein-Barr virus gp340 : CD21 Influenza virus haemagglutinin (HA) : sialic acid Penetration: e.g. fusion of virus and cell membranes (A) enveloped viruses: fusion between virus and cell membranes HIV and measles virus at cell surface Influenza virus with acidified intracellular vesicles (B) non-enveloped viruses: disruption of host cell membrane integrity, genome or core crosses into cytosol Polio virus T4 in E. coli The eclipse phase: expression of virus proteins and replication of nucleic acid. Virus particles have been disassembled so no infectious particles are present Expression of virus proteins Highly regulated: temporal and quantitative Replication of virus nucleic acid Assembly: production of new infectious particles Release: cell lysis or budding Polio The virion is 20nm and is very stable to acid pH and proteases. It is a simple mRNA molecule that is used to synthesise giant polyprotiens. It uses a complementary mRNA strand intermediate to replicate itself. Influenza Genome transcribed into mRNAs and cRNAs by virus RNA-dependent RNA polymerase. mRNAs translated to virus protein. cRNAs are copied into virus RNA (vRNA) HIV A retrovirus, (replicates via reverse transcription) Enveloped particle, 110 nm ss RNA genome (+ve), 10 kb, diploid RNA genome converted into dsDNA via reverse transcription DNA integrates into host DNA (= provirus), may remain dormant Enables vertical transmission of virus Expressed by host RNA pol II, to make HIV mRNAs, regulated by splicing Her pes Sim plex Vir us Virion: icosahedral capsid surrounded by lipid envelope, 130 nm Genome: Linear ds DNA 152 kb, ~ 80 genes Replication: in nucleus, may replicate via lytic cycle, or become latent HSV-1, cold sores; HSV-2, genital herpes Infection: via skin abrasions Lytic replication: cascade of gene expression o IE proteins: regulatory o DE proteins: replicative, e.g. DNA polymerase o Late proteins: structural o Each class requires prior expression of proteins of the previous class 3. Define the following terms as used in the description and classification of viruses: DNA virus, RNA virus, capsid type, envelope non-enveloped DNA Virus: uses DNA as its genetic material. They may be double or single stranded. RNA Virus: uses RNA as genetic material. Capsid Type: the type of outer shell the virus has. Helical capsids, Baltimore Classification icosahedral (isometric) capsids, or Group Contains enveloped. I dsDNA viruses Enveloped: Coating for virus particle. II ssDNA viruses III dsRNA viruses IV (+)ssRNA viruses V (-)ssRNA viruses VI ssRNA-RT viruses VII dsDNA-RT viruses ss: single-stranded, ds: double stranded RT: reverse transcribing