Chapter 15 Viruses I. Introduction to Viruses Virus originates from Latin word poison .

First virus discovered was tobacco mosaic disease virus (TMV) in 1890s. In 1930s: TMV was isolated and purified. By 1950s science of virology was well established. II. Characteristics of viruses They are a cellular, that is, they contain no cytoplasm or cellular organelles. They are totally dependent on a host cell for replication. (They are strict intracellular parasites) Viruses contain a single type of nucleic acid (DNA or RNA, never both) and a protein coat (capsid), sometimes enclosed by an envelope composed of lipids, proteins, and carbohydrates. Do not divide by binary fission or mitosis Viruses are usually much smaller than bacteria and are submicroscopic. Most range in size from 5 to 300 nanometers (nm), although some Paramyxoviruses can be up to 14,000nm long

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III. General Morphology An intact infectious viral particle is called a virion Helical viruses (for example, Tobacco mosaic virus) resemble long rods and their capsids are hollow cylinders surrounding the nucleic acid. Polyhedral viruses (for example, adenovirus) are many-sided. Usually the capsid is an icosahedron. , with 20 triangular faces and 12 corners. Enveloped viruses are covered by an envelope. There are enveloped helical viruses (for example, Influenzavirus) and enveloped polyhedral viruses (for example, Herpesvirus). Complex viruses have complex structures. For example, many bacteriophages have a polyhedral capsid with a helical tail attached.

1.

2. 3.

4.

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Nucleic Acid Viruses contain either DNA or RNA, never both, and the nucleic acid may be single- or double-stranded, linear or circular, or divided into several separate molecules Smallest viruses have only 4 genes, while largest have several hundred Capsid and envelope The protein coat surrounding the nucleic acid of a virus is called the capsid. The capsid is composed of subunits, capsomeres, which can be a single type of protein or several types. The capsid of some viruses is enclosed by an envelope consisting of lipids, proteins, and carbohydrates. For most viruses, is derived from host cell membranes by a process called budding Some envelopes are covered with carbohydrate-protein complexes called spikes. These glycoprotein spikes function in attaching the virus to receptors on host cells

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IV. Taxonomy of viruses Classification of viruses is based on type of nucleic acid, strategy for replication, and morphology. Nomenclature of viruses 1. Family groups ending in viridae (virus) Picornaviridae are the small RNA viruses Pico =small, rna = RNA, viridae = virus 2. Genus names end in virus Based on location they infect Enterovirus infect the enteric tract and is a subdivision of Picornaviridae 3. Species names are not italicized and not capitalized A group of viruses sharing the same genetic information and ecological niche (host). Name of the disease used as species name Poliovirus is a species of Enterovirus Subspecies are designated by a number

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V. Isolation, cultivation, and identification of viruses 1. Growth of bacteriophages in the laboratory The plaque method mixes bacteriophages with host bacteria and nutrient agar. After several viral multiplication cycles, the bacteria in the area surrounding the original virus are destroyed; the area of lysis is called a plaque. Each plaque originates with a single viral particle; the concentration of viruses is given as plaque-forming units (PFUs).

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2. Growth of animal viruses in the laboratory a. In living animals or in embryonic eggs

a. Cell cultures Cells growing in culture media in the laboratory. Primary cell lines and embryonic diploid cell lines grow for a short time in vitro. Continuous cell lines can be maintained in vitro indefinitely.

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3. Viral Identification Immunologic assays: Detect specific viral proteins or antibodies to them. Examples are: Western blotting, ELISA, Biological assays: Detect cytopathic effects (CPE) caused by viral infection of cells. o Cytopathic effects include the stopping of mitosis, lysis, formation of inclusion bodies, chromosomal changes, and transformation o Some viruses cause cytocidal effects (cell death), and others cause noncytocidal effects Plaque assays for lytic viruses Hemagglutination assay: Many viruses clump red blood cells.

Molecular assays: Assay for viral nucleic acids. o PCR (Polymerase chain reaction) o Southerns (DNA) or Northerns (RNA) Electron microscope

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VI. Viral multiplication 1. Multiplication of bacteriophages Lytic cycle: Phage causes lysis and death of host cell Lysogenic cycle: Prophage DNA incorporated in host DNA a. The lytic life cycle T-even bacteriophages that infect E. coli The virus life cycle can be divided into five stages: 1. Attachment phase of the lytic cycle, sites on the phage's tail fibers attach to com-plementary receptor sites on the bacterial cell. 2. Penetration phase, phage lysozyme opens a portion of the bacterial cell wall, the tail sheath contracts to force the tail core through the cell wall, and phage DNA enters the bacterial cell. The capsid remains outside. 3. Biosynthesis, transcription of phage DNA produces mRNA coding for proteins necessary for phage multiplication. Phage DNA is replicated, and capsid proteins are produced. 4. Maturation, phage DNA and capsids assemble into complete viruses. 5. Release, phage lysozyme breaks down the bacterial cell wall, and the multiplied phages are released

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The one-step growth curve of virus replication Following adsorption, the infectivity of the virus particles disappears, a phenomenon called eclipse. This is due to the uncoating of the virus particles. During the latent period, replication of viral nucleic acid and protein occurs. The maturation period and release follow The time from phage adsorption to release is called burst time (20 to 40 minutes). Burst size, the number of newly synthesized phages produced from a single infected cell, ranges from 50 to 200.

b. The Lysogenic Life Cycle Bacteriophages capable of a lysogenic life cycle are termed temperate phages. Phage lambda, infects E. coli 1. Attachment and Penetration: Virus tail binds to specific receptors on the cell surface and injects genetic material (DNA) into cell. 2. Circularization: Phage DNA circularizes and enters either lytic or lysogenic cycle. Lysogenic cycle 3. Integration: Phage DNA integrates with bacterial chromosome and becomes a prophage . Prophage remains latent. 4. Excision: Prophage DNA is removed due to a stimulus (e.g.: chemicals, UV radiation) and initiates a lytic cycle.
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Results of lysogeny Lysogenic cells are immune to reinfection by the same phage Host cell may exhibit new properties C. diphtheria - Toxin production requires phage virus. 2. Multiplication of animal viruses 1. Animal viruses attach to the plasma membrane of the host cell. 2. Penetration occurs by endocytosis or fusion

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3. Animal viruses are uncoated by viral or host cell enzymes. 4. Replication of NA and protein synthesis Animal virusesgrouped by type of nucleic acid There are three possible patterns of viral genome replication: a. DNA DNA. If viral DNA is double-stranded, DNA replication resembles that of cellular DNA, and the virus uses DNA polymerase produced by the host.

b. RNA RNA. Since host cells lack the enzyme to copy RNA, most RNA viruses contain a gene that codes for RNA replicase, an enzyme that uses viral RNA as a template to produce complementary RNA. Picornaviridae + strand RNA acts as mRNA and directs the synthesis
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of RNA-dependent RNA polymerase. Rhabdoviridae - strand RNA is a template for viral RNA-dependent RNA polymerase, which transcribes mRNA. Reoviridae (double capsid) are digested in host-cell cytoplasm to release double-stranded RNA for viral biosynthesis. c. RNA DNA RNA. Some RNA viruses encode reverse transcriptase, an enzyme that transcribes DNA from an RNA template Retrovirus = (Retro = backward) RNA virus that uses reverse transcriptase to transcribe DNA from the viral RNA genome. Reverse transcriptase is a type of DNA polymerase that transcribes DNA from an RNA template. HIV (human immunodeficiency virus), the virus that causes AIDS (acquired immunodeficiency syndrome) is a retrovirus.

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5. After maturation, viruses are released. One method of release (and envelope formation) is budding. Nonenveloped viruses are released through ruptures in the host cell membrane. VII. Viruses and Cancer The earliest relationship between cancer and viruses was demonstrated in the early 1900s, when chicken leukemia and chicken sarcoma (cancer of connective tissue) were transferred to healthy animals by cell-free filtrates. Several viral genes have been identified as oncogenes. Oncogenes = Genes found in viruses or as part of the normal eukaryotic genome, that trigger transformation of a cell to a cancerous state. Code for cellular growth factors or for proteins involved in the function of growth factors. Some tumor viruses transform cells by activating cellular oncogenes. Viruses capable of producing tumors are called oncogenic viruses. The genetic material of oncogenic viruses becomes integrated into the host cell's DNA. Transformed cells lose contact inhibition, exhibit chromosomal abnormalities, and can produce tumors when injected into susceptible animals. Certain viruses are implicated in human cancers:

The retroviruses ability to produce tumors is related to the production of reverse transcriptase. The DNA synthesized from the viral RNA becomes incorporated as a provirus into the host cell's
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DNA. A provirus can remain latent, can produce viruses, or can transform the host cell. Prions Prions are infectious proteins first discovered in the 1980s. Prion diseases, such as Creutzfeldt-Jakob disease (CJD) and mad cow disease, all involve degeneration of brain tissue. Prion diseases are due to an altered protein; the cause can be a mutation in the normal gene for PrP or contact with an altered protein (PrpSc). Viroids Infectious pieces of RNA that cause some plant diseases, such as potatoes, and tomatoes X. Antiviral therapeutic agents Amantadine Influenza Inhibits uncoating Acyclovir Herpes simplex Inhibits DNA polymerase Azidothymidime HIV Inhibits reverse transcriptase Interferon Cytomegalovirus Inhibits protein synthesis

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Viral vaccines First vaccine was used by Jenner (1798) against smallpox and contained live vaccinia (cowpox) virus. 1. Live attenuated vaccines: Mutant viral strains produce an asymptomatic infection in host. Examples: Polio (oral, Sabin vaccine), measles, yellow fever, mumps, rubella, and chickenpox. Advantages: Better immune response Disadvantages: May cause disease due to contamination, genetic instability 2. Killed or inactivated vaccines formalin.
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Examples: Rabies, and influenza A & B. Advantages: Immunization with little or no risk of infection. Disadvantages: Less effective immune response 3. Recombinant vaccines: Viral subunits are produced by genetically engineered cells. Example: Hepatitis B Advantages: Little or no risk of infection. Disadvantages: Less effective immune response.

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