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Virus structure and replication

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VIRUS STRUCTURE & REPLICATION

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Virus structure and replication

  1. 1. VIRUS STRUCTURE & REPLICATION Dr.T.V.Rao MD Dr.T.V.Rao MD 1
  2. 2. What are viruses?What are viruses? • Small obligate intracellular parasitesSmall obligate intracellular parasites • VirionVirion – Complete virus particle : nucleic acid + protein coat, which may be surrounded by an envelope – It is the form in which the virus moves between cells or hosts • Viral GenomeViral Genome – EITHER RNA or DNA genome surrounded by a protective virus-coded protein coat (Capsid) • Propagation depends on specialized host cells supplying the machinery for replication, metabolism and biosynthesis Dr.T.V.Rao MD 2
  3. 3. Configuration of VirusConfiguration of Virus • The DNA or RNA genome may be : – ss – single stranded or – ds – double stranded • Genomes may be either: – (+) sense: Positive-sense viral RNA is identical to viral mRNA and thus can be immediately translated into protein by the host cell. OR – (-) sense: Negative-sense viral RNA is complementary to mRNA and thus must be converted to positive-sense RNA by an RNA polymerase before translation. • Retroviruses? Dr.T.V.Rao MD 3
  4. 4. Viral Structure - Overview Fig 1. Schematic overview of the structure of animal viruses ** does not exist in all viruses Nucleic acid Capsid Nucleocapsid Envelope protein Membrane protein Viral envelope** Spike protein Dr.T.V.Rao MD 4
  5. 5. Bacteriophages http://micro.magnet.fsu.edu/cells/virus.html Dr.T.V.Rao MD 5
  6. 6. Viroids & PrionsViroids & Prions • Viroids – ss RNA genome and the smallest known pathogens. – Affects plants • Prions – Infectious particles that are entirely protein. – No nucleic acid – Highly heat resistant – Animal disease that affects nervous tissue – Affects nervous tissue and results in • Bovine spongiform encephalitis (BSE) “mad cow disease”, • Scrapie in sheep • kuru & Creutzfeldt-Jakob Disease (CJD) in humans Dr.T.V.Rao MD 6
  7. 7. Viral Structure • Varies in size, shape and symmetry • VIP for classification • 3 types of capsid symmetry: – Cubic (icosahedral) • Has 20 faces, each an equilateral triangle. Eg. adenovirus – Helical • Protein binds around DNA/RNA in a helical fashion eg. Coronavirus – Complex • Is neither cubic nor helical eg. poxvirus Dr.T.V.Rao MD 7
  8. 8. Viral Structure Figure 1 An array of viruses. (a) The helical virus of rabies. (b) The segmented helical virus of influenza. (c) A bacteriophage with an icosahedral head and helical tail. (d) An enveloped icosahedral herpes simplex virus. (e) The unenveloped polio virus. (f) The icosahedral HIV with spikes on its envelope. Dr.T.V.Rao MD 8
  9. 9. Dr.T.V.Rao MD 9
  10. 10. DefinitionsDefinitions • Bacteriophage – Virus that infects prokaryotic (bacterial) cells. • Nucleocapsid: –viral nucleic acid + the protein coat that encloses it. –Represents the packaged form of the viral genome. Dr.T.V.Rao MD 10
  11. 11. Viral Replication • Viruses are intracellular obligate parasites which means that they cannot replicate or express their genes without the help of a living cell. A single virus particle (Virion) is in and of itself essentially inert. It lacks needed components that cells have to reproduce. When a virus infects a cell, it marshals the cell's ribosomes, enzymes and much of the cellular machinery to replicate. Dr.T.V.Rao MD 11
  12. 12. Progress of Viral Multiplication • Once a virus infects its host and the viral progeny components are produced by the host's cellular machinery, the assembly of the viral capsid is a non- enzymatic process. It is usually spontaneous. Viruses typically can only infect a limited number of hosts (also known as host range). The "lock and key" mechanism is the most common explanation for this range. Certain proteins on the virus particle must fit certain receptor sites on the particular host's cell surface. Dr.T.V.Rao MD 12
  13. 13. • When a virus infects a cell, nucleic acid must be uncoated and gain access to metabolic machinery of cell. • Virus life cycle is characterized by: – attachment – penetration, with entry of nucleic acid into cell – early expression of virus genes (either directly by translation, if virus contains "+" RNA, or indirectly after transcription and then translation) – replication of virus nucleic acid – synthesis of new Virion components – packaging and assembly of new virions – exit from cell Viral Replication Dr.T.V.Rao MD 13
  14. 14. Pricipliple of Replication  Replication of the genome is governed by the principle of complementarity  requires a strand with complementary base sequence be synthesized  serve as template  synthesis of actual viral genome  Late synthesis  viral structural proteins for the capsid are also synthesized almost simultaneously Dr.T.V.Rao MD 14
  15. 15. IMPORTANT FACTS ABOUT VIRUSIMPORTANT FACTS ABOUT VIRUS REPLICATIONREPLICATION  Viruses multiply only in living cells  Host cell provides the energy & machinery for the synthesis of viral proteins & nucleic acids  Viral genome must be able to produce mRNA  host cell protein-synthesizing machinery may be able to synthesize viral proteins Dr.T.V.Rao MD 15
  16. 16. Stages in Multiplication  Soon after interaction with the host cell  Virion is disrupted  infectivity is lost  ECLIPSE PERIOD  The yield of infectious virus per cell ranges from moderate numbers to > 100,000 particles  The duration of replication cycle varies from 6-8 hrs to > 40 hrs Dr.T.V.Rao MD 16
  17. 17. All infections are not Productive  PRODUCTIVE INFECTIONS > occur in permissive cells  infectious virus  ABORTIVE INFECTIONS > no infectious virus produce because : a. cell is non-permissive b. virus may be defective DEFECTIVE VIRUS – lack certain genes for replication & requires the help of another virus (dependo or helper virus) Dr.T.V.Rao MD 17
  18. 18. LATENT INFECTIONSLATENT INFECTIONS  LATENT INFECTIONSLATENT INFECTIONS a. viral genome persists within the host cell b. expression of no or few viral genes c. survival of the infected cell Dr.T.V.Rao MD 18
  19. 19. Pricipliple of Replication  Replication of the genome is governed by the principle of complementarity  requires a strand with complementary base sequence be synthesized  serve as template  synthesis of actual viral genome  Late synthesis  viral structural proteins for the capsid are also synthesized almost simultaneously Dr.T.V.Rao MD 19
  20. 20. STEPS IN VIRAL REPLICATIONSTEPS IN VIRAL REPLICATION 1. Attachment & adsorption 2. Penetration 3. Uncoating 4. Early viral mRNA synthesis 5. Early viral protein synthesis Dr.T.V.Rao MD 20
  21. 21. STEPS IN VIRAL REPLICATION 6. Viral genome replication 7. Late viral mRNA synthesis 8. Late viral protein synthesis 9. Assembly 10. Release Dr.T.V.Rao MD 21
  22. 22. Early steps in MultiplicationEarly steps in Multiplication • Attachment – specific binding of a Virion protein (the anti-receptor) to a constituent of the cell surface (the receptor) • e.g. hemagglutinin of influenza virus • some complex viruses (HSV) may have more than one species of anti-receptor molecule • Penetration – energy-dependent step – occurs almost instantaneously after attachmentDr.T.V.Rao MD 22
  23. 23. ASSEMBLY & RELEASE  It is thru budding that the virus acquire its envelope  Budding process begin when virus-specific proteins enter the cell membrane at specific sites  Herpesviruses  nuclear membrane Poxviruses  inclusion bodies Coronaviruses  endoplasmic reticulum Dr.T.V.Rao MD 23
  24. 24. ASSEMBLY & RELEASE  Viral nucleocapsid then interacts with the specific membrane site mediated by matrix protein  Cell membrane evaginates at that site  an enveloped particle bud off from the membrane Dr.T.V.Rao MD 24
  25. 25. ATTACHMENT & ADSORPTION  Reversible step  Does not require energy  2 requirements: 1. recognition & attachment to specific host receptor  determines the host range of the virus Dr.T.V.Rao MD 25
  26. 26. Penetration and Fusion Dr.T.V.Rao MD 26
  27. 27. Mechanism differ the various types > Rhinovirus – ICAM-1 > HIV – CD4 + T cells > Rabies virus – acetylcholine receptors > HSV-1 – fibroblast growth factor receptor 2. appropriate ph & ionic concentration  both the host cell & viral particle are negatively charged at ph 7  require counter-ion  Magnesium ion Dr.T.V.Rao MD 27
  28. 28. PENETRATION  Refers to the entry of the viral particle into the cytoplasm of the host cell  Temperature-dependent step (37 C)  Carried out through: a. receptor-mediated endocytosis b. direct penetration (viropexis) c. cell fusion Dr.T.V.Rao MD 28
  29. 29. UNCOATINGUNCOATING • Refers to the physical separation of the viral capsid from the viral genome • Considered an obligatory step in viral replication  makes the viral genome accessible for transcription • Favored by low pH Dr.T.V.Rao MD 29
  30. 30. Assembly and ReleaseAssembly and Release • Components of capsid synthesis directed by late genes • Assembly of enveloped viruses needs interaction with plasma membrane which has been modified • Final stage of infection • Enveloped viruses released gradually by budding or exocytosis • Naked viruses accumulate in cytoplasm and released Dr.T.V.Rao MD 30
  31. 31. INFECTIOUS NUCLEIC ACID  Purified viral nucleic acid (without any protein) that can carry out the entire viral growth cycle  complete viral particles  Can bypass the host range specificity provided by the viral protein-host cell receptor interaction e.g. Poliovirus Dr.T.V.Rao MD 31
  32. 32. GENE EXPRESSION & GENOMEGENE EXPRESSION & GENOME REPLICATIONREPLICATION  1st step in gene expression  mRNA synthesis  DNA viruses Except Poxviruses replicate in the nucleus  use host cell DNA-dependent RNA polymerase  synthesize mRNA Dr.T.V.Rao MD 32
  33. 33. Multiply in  Poxviruses replicate in the cytoplasm no access to host cell RNA polymerase  carry their own RNA polymerase  RNA viruses replicate in the cytoplasm EXCEPT Influenza virus & Retrovirus Dr.T.V.Rao MD 33
  34. 34. ASSEMBLY & RELEASE  Progeny particles assembled by packaging the viral nucleic acid within the capsid proteins  2 processes of Release: 1. Rupture or lysis of cell membrane 2. Budding through the outer membrane Dr.T.V.Rao MD 34
  35. 35. Differences in Single Stranded and Double Stranded Virus Double-stranded RNA  cell has no enzyme to transcribe RNA into mRNA  virus carries its own polymerase Single-stranded RNA (+ polarity) e.g. Retrovirus  carries RNA-dependent DNA polymerase  ds DNA  transcribed to mRNA by host cell polymerase Dr.T.V.Rao MD 35
  36. 36. Synthesis of RNA  Once m RNA is synthesized  translated into viral proteins (enzymes & early proteins for the genome) by host ribosomes  The most important of the early proteins is the Polymerase  synthesize many copies of viral genetic material for the progeny virus particle Dr.T.V.Rao MD 36
  37. 37. Synthesis of PapovaVirus Dr.T.V.Rao MD 37
  38. 38. RNARNA GenomeGenome PolarityPolarity VirionVirion Polyme-Polyme- raserase SourceSource OfOf mRNAmRNA InfectivityInfectivity OfOf GenomeGenome PrototypePrototype SS, nonSS, non segmentedsegmented ++ NoNo GenomeGenome YesYes PoliovirusPoliovirus SSSS Nonseg-Nonseg- mentedmented -- YesYes Transcrip-Transcrip- tiontion NoNo MeaslesMeasles RabiesRabies segmentedsegmented -- YesYes Transcrip-Transcrip- tiontion NoNo InfluenzaInfluenza DSDS segmentedsegmented +/-+/- YesYes Transcrip-Transcrip- tiontion NoNo ReovirusReovirus SS diploidSS diploid ++ YesYes Transcrip-Transcrip- tiontion NoNo HLTVHLTV HIVHIV Dr.T.V.Rao MD 38
  39. 39. Dr.T.V.Rao MD 39
  40. 40. To Learn More about Microbiology and Infectious Diseases Join Dr.T.V.Rao MD 40
  41. 41. Microbiology at your Finger Tips • To Learn More about Microbiology and Infectious Diseases Join • Rao’s Microbiology and • Rao’s Infection careRao’s Infection care onon FACEBOOK Dr.T.V.Rao MD 41
  42. 42. • Programme Created by Dr.T.V.Rao MD for Medical and Paramedical Students in the Developing World • Email • doctortvrao@gmail.comdoctortvrao@gmail.com Dr.T.V.Rao MD 42

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