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Chapter 13. Viruses, Viroids and Prions. Adolf Mayer, 1886 tobacco mosaic disease (TMD) transmissible Dimitri Iwanowski, 1892 Filtered sap still caused TMD contagious fluid or filterable agent Walter Reed,1901 Yellow Fever. Felix d’Herelle, 1917 Bacteriophage Suggested phage therapy

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Chapter 13

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    1. Chapter 13 Viruses, Viroids and Prions

    2. Adolf Mayer, 1886 • tobacco mosaic disease (TMD) transmissible • Dimitri Iwanowski, 1892 • Filtered sap still caused TMD • contagious fluid or filterable agent • Walter Reed,1901 • Yellow Fever

    3. Felix d’Herelle, 1917 • Bacteriophage • Suggested phage therapy • 1930’s, term virus introduced and electron microscopy invented • Wendell Stanley, 1935 • Isolated tobacco mosaic virus

    4. [INSERT FIGURE 13.4]

    5. Viral Features • Obligate intracellular pathogen • Host range • highly specific or generalists • DNA or RNA • Protein coat • Surface proteins highly susceptible to mutations • Cause synthesis of specialized structures to transfer viral particles to other cells

    6. Virion • complete, fully developed, infectious viral particle • Living or non-living entities?????

    7. Viral Classification • Oldest system based on symptomology • International Committee on Taxonomy of Viruses (ICTV), 1966 • Describe viruses as elementary bio-systems • Classified into orders, families, genera and species • Over 1,500 officially recognized species

    8. 3 primary means of classification: Nucleic acid Replication strategy Morphology of protein coat (capsid)

    9. Helical Viruses Polyhedral Viruses

    10. Viral Envelope • Acquired from host cell • Phospholipids and proteins • Some glycoproteins are virally coded spikes • Often play role in host recognition

    11. Critical Swine Flu prevention tip: Don't DO this!

    12. Complex Viruses

    13. [INSERT FIGURE 13.5]

    14. Viral Taxonomy • Order –virales • Family –viridae • Genus – virus • Species • Common names • Subspecies designated by a number

    15. Viral Taxonomy • Retroviridae– family • Lentivirus – genus • Human Immunodeficiency Virus– species • Herpesviridae • Simplexvirus • Human herpesvirus 1, HHV 2, HHV 3

    16. Isolation and Cultivation of Viruses • Viruses must be grown in living cells • Cytopathic effects

    17. Animal viruses may be grown in living animals or in embryonated eggs

    18. Animal & plants viruses may be grown in cell cultures • Primary cell lines • Continuous cell lines (transformed cells )

    19. Virus Identification • Serological tests • Detect antibodies against viruses in a patient • Nucleic acids • RFLPs • PCR

    20. Lytic cycle Phage causes lysis and death of host cell Lysogenic cycle Temperate phages incorporate DNA into host DNA (prophage) Multiplication of Bacteriophages

    21. Lytic Cycle • Attachment Phage attaches by tail fibers to host cell • Penetration Phage lysozyme opens cell wall, tail sheath contracts to force tail core and DNA into cell • Biosynthesis Production of phage DNA and proteins • Maturation Assembly of newly synthesized phage particles • Release Phage lysozyme breaks cell wall

    22. [INSERT FIGURE 13.8]

    23. One-step Growth Curve

    24. Lysogenic Cycle

    25. 3 Important results of lysogeny • Immunity to re-infection • Phage conversion • Specialized transduction

    26. Specialized Transduction gal gene Bacterial DNA Prophage 1 Prophage exists in galactose-using host (containing the gal gene). Galactose-positive donor cell gal gene 2 Phage genome excises, carrying with it the adjacent gal gene from the host. 3 Phage matures and cell lyses, releasing phage carrying gal gene. gal gene 4 Phage infects a cell that cannot utilize galactose (lacking gal gene). Galactose-negative recipient cell 5 Along with the prophage, the bacterial gal gene becomes integrated into the new host’s DNA. 6 Lysogenic cell can now metabolize galactose. Galactose-positive recombinant cell

    27. Multiplication of Animal viruses • Attachment Viruses attaches to cell membrane • Penetration By endocytosis or fusion • Uncoating By viral or host enzymes • Biosynthesis Production of nucleic acid and proteins • Maturation Nucleic acid and capsid proteins assemble • Release By budding (enveloped viruses) or rupture

    28. Attachment of animal viruses • Chemical attraction • No tails or tail fibers • Glycoprotein spikes or other attachment molecules

    29. Replication of Animal Viruses • Biosynthesis • Each virus requires different strategy depending on its nucleic acid • DNA viruses often enter the nucleus • RNA viruses typically replicate in cytoplasm • Must consider: • What serves as template for replication and how mRNA is transcribed

    30. Multiplication of DNA Virus Papovavirus 1 Virion attaches to host cell 7 Virions are released Host cell DNA Capsid 2 DNA Virion penetrates cell and its DNA is uncoated Cytoplasm 6 Virions mature Capsid proteins mRNA 5 Late translation; capsid proteins are synthesized 3 Viral DNA penetrates host nucleus Early transcription and translation; enzymes are synthesized 4 Late transcription; DNA is replicated

    31. Multiplication of RNA Virus [INSERT FIGURE 13.13]

    32. Multiplication of a Retrovirus Capsid Reverse transcriptase DNA Virus Two identical + stands of RNA 1 Retrovirus penetrates host cell. Host cell DNA of one of the host cell’s chromosomes 5 Mature retrovirus leaves host cell, acquiring an envelope as it buds out. Reverse transcriptase 2 Virion penetrates cell and its DNA is uncoated Viral RNA Identical strands of RNA 4 Transcription of the provirus may also occur, producing RNA for new retrovirus genomes and RNA that codes for the retrovirus capsid and envelope proteins. Viral proteins RNA 3 The new viral DNA is tranported into the host cell’s nucleus and integrated as a provirus. The provirus may divide indefinitely with the host cell DNA. Provirus

    33. Assembly and release of animal viruses • Most DNA viruses assemble in and are released from nucleus into cytosol • Most RNA viruses develop solely in cytoplasm • Enveloped viruses cause persistent infections • Naked viruses are released by exocytosis or may cause lysis and death of host cell

    34. Viruses and Cancer • Oncogenes transform normal cells into tumor cells • Activated by mutagenic chemicals, radiation, viruses • Causes increased growth, loss of contact inhibition • Cells tend to be misshapen and exhibit chromosomal abnormalities

    35. Oncoviruses • Viral DNA integrated into host DNA • Induces tumors

    36. Oncogenic Viruses • Oncogenic DNA Viruses • Adenoviridae • Herpesviridae • Poxviridae • Papovaviridae • Hepadnaviridae • Oncogenic RNA viruses • Retroviridae • DNA • HTLV 1 • HTLV 2

    37. Latent Viral Infections • Virus remains dormant in asymptomatic host cell for long periods • Cold sores, shingles • Persistent Viral Infections • Disease progresses slowly over a long period, generally fatal • Subacute sclerosing panencephalitis (measles virus)

    38. Plant Viruses • Plant viruses enter through wounds or via biting insects • May be transmitted in pollen • Viroids • infectious naked RNA Potato Spindle Tuber Viroid

    39. Prions • Proteinaceous infectious particle • Inherited and transmissible diseases • Spongiform encephalopathies • Sheep scrapie, Creutzfeldt-Jakob disease, Gerstmann-Sträussler-Scheinker syndrome, fatal familial insomnia, mad cow disease

    40. PrPC, normal cellular prion proteinPrPSc, scrapie protein

    41. [INSERT FIGURE 13.23] scrapie proteinsaccumulate in brain cells forming large vacuoles