Lecture PowerPoint to accompany Foundations in Microbiology Sixth Edition Talaro Chapter 6 An Introduction to the Viruses Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. 2 General Structure of Viruses • Size range – – most <0.2 μm; requires electron microscope • Virion – fully formed virus able to establish an infection 3 4 General Structure of Viruses • Capsids – All viruses have capsids - protein coats that enclose and protect their nucleic acid. – Each capsid is constructed from identical subunits called capsomers made of protein. – The capsid together with the nucleic acid are nucleoscapsid. – Some viruses have an external covering called envelope; those lacking an envelope are naked 5 6 General Structure of Viruses • Two structural types: – helical - continuous helix of capsomers forming a cylindrical nucleocapsid – icosahedral - 20-sided with 12 corners – vary in the number of capsomers – Each capsomer may be made of 1 or several proteins. – Some are enveloped. 7 8 9 General Structure of Viruses • Viral envelope – mostly animal viruses – acquired when the virus leaves the host cell – exposed proteins on the outside of the envelope, called spikes, essential for attachment of the virus to the host cell 10 Functions of Capsid/Envelope • Protects the nucleic acid when the virion is outside the host cell • Helps to bind the virion to a cell surface and assists the penetration of the viral DNA or RNA into a suitable host cell 11 General Structure of Viruses • Complex viruses: atypical viruses – Poxviruses lack a typical capsid and are covered by a dense layer of lipoproteins. – Some bacteriophages have a polyhedral nucleocapsid along with a helical tail and attachment fibers. 12 13 Nucleic acids • Viral genome – either DNA or RNA but never both • Carries genes necessary to invade host cell and redirect cell’s activity to make new viruses • Number of genes varies for each type of virus – few to hundreds 14 Nucleic Acids • DNA viruses – usually double stranded (ds) but may be single stranded (ss) – circular or linear • RNA viruses – usually single stranded, may be double stranded, may be segmented into separate RNA pieces – ssRNA genomes ready for immediate translation are positive-sense RNA. – ssRNA genomes that must be converted into proper form are negative-sense RNA. 15 General Structure • Pre-formed enzymes may be present. – polymerases – DNA or RNA – replicases – copy DNA – reverse transcriptase –synthesis of DNA from RNA (AIDS virus) 16 How Viruses are Classified • Main criteria presently used are structure, chemical composition, and genetic makeup. • No taxa above Family (no kingdom, phylum, etc.) • Currently recognized: 3 orders, 63 families, and 263 genera of viruses • Family name ends in -viridae, i.e.Herpesviridae • Genus name ends in -virus, Simplexvirus • Herpes simplex virus I (HSV-I) 17 Modes of Viral Multiplication General phases in animal virus multiplication cycle: 1. Adsorption - binding of virus to specific molecule on host cell 2. Penetration - genome enters host cell 3. Uncoating – the viral nucleic acid is released from the capsid 4. Synthesis – viral components are produced 5. Assembly – new viral particles are constructed 6. Release – assembled viruses are released by budding (exocytosis) or cell lysis 18 19 Adsorption and Host Range • Virus coincidentally collides with a susceptible host cell and adsorbs specifically to receptor sites on the cell membrane • Spectrum of cells a virus can infect – host range – hepatitis B – human liver cells – poliovirus – primate intestinal and nerve cells – rabies – various cells of many mammals 20 21 Penetration/Uncoating • Flexible cell membrane is penetrated by the whole virus or its nucleic acid by: – endocytosis – entire virus is engulfed and enclosed in a vacuole or vesicle – fusion – envelope merges directly with membrane resulting in nucleocapsid’s entry into cytoplasm 22 23 Replication and Protein Production • Varies depending on whether the virus is a DNA or RNA virus • DNA viruses generally are replicated and assembled in the nucleus. • RNA viruses generally are replicated and assembled in the cytoplasm. – Positive-sense RNA contain the message for translation. – Negative-sense RNA must be converted into positive-sense message. 24 Release • Assembled viruses leave host cell in one of two ways: – budding – exocytosis; nucleocapsid binds to membrane which pinches off and sheds the viruses gradually; cell is not immediately destroyed – lysis – nonenveloped and complex viruses released when cell dies and ruptures • Number of viruses released is variable – 3,000-4,000 released by poxvirus – >100,000 released by poliovirus 25 26 Damage to Host Cell Cytopathic effects - virus-induced damage to cells 1. Changes in size & shape 2. Cytoplasmic inclusion bodies 3. Nuclear inclusion bodies 4. Cells fuse to form multinucleated cells. 5. Cell lysis 6. Alter DNA 7. Transform cells into cancerous cells 27 Persistent Infections • Persistent infections - cell harbors the virus and is not immediately lysed • Can last weeks or host’s lifetime; several can periodically reactivate – chronic latent state – measles virus – may remain hidden in brain cells for many years – herpes simplex virus – cold sores and genital herpes – herpes zoster virus – chickenpox and shingles 28 • Some animal viruses enter host cell and permanently alter its genetic material resulting in cancer – transformation of the cell. • Transformed cells have increased rate of growth, alterations in chromosomes, and capacity to divide for indefinite time periods resulting in tumors. • Mammalian viruses capable of initiating tumors are called oncoviruses. – Papillomavirus – cervical cancer – Epstein-Barr virus – Burkitt’s lymphoma 29 Multiplication Cycle in Bacteriophages • Bacteriophages – bacterial viruses (phages) • Most widely studied are those that infect Escherichia coli – complex structure, DNA • Multiplication goes through similar stages as animal viruses. • Only the nucleic acid enters the cytoplasm uncoating is not necessary. • Release is a result of cell lysis induced by viral enzymes and accumulation of viruses - lytic cycle. 30 6 Steps in Phage Replication 1. Adsorption – binding of virus to specific molecule on host cell 2. Penetration –genome enters host cell 3. Replication – viral components produced 4. Assembly - viral components assembled 5. Maturation – completion of viral formation 6. Release – viruses leave cell to infect other cells 32 Lysogeny: The Silent Virus Infection • Not all phages complete the lytic cycle. • Some DNA phages, called temperate phages, undergo adsorption and penetration but don’t replicate. • The viral genome inserts into bacterial genome and becomes an inactive prophage - the cell is not lysed. • Prophage is retained and copied during normal cell division resulting in the transfer of temperate phage genome to all host cell progeny – lysogeny. • Induction can occur resulting in activation of lysogenic prophage followed by viral replication and cell lysis. 33 34 Lysogeny • Lysogeny results in the spread of the virus without killing the host cell. • Phage genes in the bacterial chromosome can cause the production of toxins or enzymes that cause pathology – lysogenic conversion. – Corynebacterium diphtheriae – Vibrio cholerae – Clostridium botulinum 35 36 Techniques in Cultivating and Identifying Animal Viruses • Obligate intracellular parasites that require appropriate cells to replicate • Methods used: – cell (tissue) cultures – cultured cells grow in sheets that support viral replication and permit observation for cytopathic effect – bird embryos – incubating egg is an ideal system; virus is injected through the shell – live animal inoculation – occasionally used when necessary 37 Prions and Other Infectious Particles Prions - misfolded proteins, contain no nucleic acid – – cause transmissible spongiform encephalopathies – fatal neurodegenerative diseases common in animals: • • • • • scrapie in sheep & goats bovine spongiform encephalopathies (BSE), aka mad cow disease wasting disease humans – Creutzfeldt-Jakob Syndrome (CJS) Extremely resistant to usual sterilization techniques 38 Other noncellular infectious agents • Satellite viruses – dependent on other viruses for replication – adeno-associated virus – replicate only in cells infected with adenovirus – delta agent – naked strand of RNA expressed only in the presence of hepatitis B virus • Viroids - short pieces of RNA, no protein coat; only been identified in plants, so far 39 Detection and Treatment of Animal Viral Infections • More difficult than other agents • Consider overall clinical picture • Take appropriate sample – Infect cell culture- look for characteristic cytopathic effects – Screen for parts of the virus – Screen for immune response to virus (antibodies) • Antiviral drugs can cause serious side effects 40