Chapter 13 Characterizing and Classifying Viruses, Viroids, and Prions © 2012 Pearson Education Inc. Lecture prepared by Mindy Miller-Kittrell North Carolina State University Characteristics of Viruses • Virus – Minuscule, acellular infectious agent having either DNA or RNA – Causes many infections of humans, animals, plants, and bacteria – Causes most of the diseases that plague the industrialized world © 2012 Pearson Education Inc. Characteristics of Viruses • Cannot carry out any metabolic pathway • Neither grow nor respond to the environment • Cannot reproduce independently • Recruit the cell’s metabolic pathways to increase their numbers • No cytoplasmic membrane, cytosol, organelles (with one exception) • Have extracellular and intracellular state © 2012 Pearson Education Inc. Characteristics of Viruses • Extracellular State – Called virion – Protein coat (capsid) surrounding nucleic acid – Nucleic acid and capsid also called nucleocapsid – Some have phospholipid envelope – Outermost layer provides protection and recognition sites for host cells • Intracellular State – Capsid removed – Virus exists as nucleic acid © 2012 Pearson Education Inc. Figure 13.1 Virions-overview Characteristics of Viruses • Genetic Material of Viruses – Show more variety in nature of their genomes than do cells – Primary way scientists categorize and classify viruses – May be DNA or RNA, but never both – dsDNA, ssDNA, dsRNA, ssRNA – Linear and segmented or single and circular – Much smaller than genomes of cells © 2012 Pearson Education Inc. Figure 13.2 The relative sizes of genomes Partial genome of E. coli Viral genome Characteristics of Viruses • Hosts of Viruses – Most viruses infect only particular host’s cells – Affinity of viral surface proteins for proteins on host cell – May be so specific they infect only particular kind of cell in a particular host – Generalists – infect many kinds of cells in many different hosts © 2012 Pearson Education Inc. Figure 13.3 Hosts of viral infections-overview Figure 13.4 Sizes of selected virions E. coli (bacterium) (1000 nm 3000 nm) Red blood cell (10,000 nm in diameter) Bacterial ribosomes (25 nm) Poliovirus (30 nm) Bacteriophage MS2 (24 nm) Smallpox virus (200 nm 300 nm) Bacteriophage T4 (50 nm 225 nm) Tobacco mosaic virus (15 nm 300 nm) Characteristics of Viruses • Capsid Morphology – Capsids – Provide protection for viral nucleic acid – Means of attachment to host’s cells – Composed of proteinaceous subunits called capsomeres – Capsomere made of single or multiple types of proteins © 2012 Pearson Education Inc. Characteristics of Viruses • Viral Shapes – Three basic shapes – Helical – Polyhedral – Complex © 2012 Pearson Education Inc. Figure 13.5 The shapes of virions-overview Figure 13.6 Bacteriophage T4-overview Characteristics of Viruses • The Viral Envelope – Acquired from host cell during viral replication or release – Envelope is portion of membrane system of host – Composed of phospholipid bilayer and proteins – Some proteins are virally coded glycoproteins (spikes) – Envelope’s proteins and glycoproteins often play role in host recognition © 2012 Pearson Education Inc. Figure 13.7 Enveloped virion-overview Table 13.2 Families of Human Viruses Viral Replication • Dependent on hosts’ organelles and enzymes to produce new virions • Lytic replication – Replication cycle usually results in death and lysis of host cell • Stages of lytic replication cycle – – – – – Attachment Entry Synthesis Assembly Release © 2012 Pearson Education Inc. Viral Replication ANIMATION Viral Replication: Overview © 2012 Pearson Education Inc. Figure 13.8 The lytic replication cycle in bacteriophages-overview Attachment Bacteriophage genome Entry Tail sheath Outer membrane Peptidoglycan Cytoplasmic membrane Bacterial chromosome Entry Attachment Phage DNA Lytic replication cycle of bacteriophage Bacterial chromosome degraded Release Synthesis Phage proteins Assembly Assembly Base Tail Sheath DNA Capsid Mature head Tail fibers Mature virion Number of infective virions in medium (log scale) Figure 13.9 Pattern of virion abundance in lytic cycle Burst size Entry Synthesis and assembly Time (minutes) Attachment Burst time Viral Replication ANIMATION Viral Replication: Virulent Bacteriophages © 2012 Pearson Education Inc. Viral Replication • Lysogeny – Modified replication cycle – Infected host cells grow and reproduce normally for generations before they lyse – Temperate phages – Prophages – inactive phages – Lysogenic conversion results when phages carry genes that alter phenotype of a bacterium © 2012 Pearson Education Inc. Figure 13.10 Bacteriophage lambda Figure 13.11 The lysogenic replication cycle in bacteriophages: phage lambda and E. coli Attachment Prophage in chromosome Entry Lambda phage Lytic cycle Lysogeny Synthesis Release Replication of chromosome and virus; cell division Assembly Induction Further replications and cell divisions Viral Replication ANIMATION Viral Replication: Temperate Bacteriophages © 2012 Pearson Education Inc. Viral Replication • Replication of Animal Viruses – Same basic replication pathway as bacteriophages – Differences result from – Presence of envelope around some viruses – Eukaryotic nature of animal cells – Lack of cell wall in animal cells © 2012 Pearson Education Inc. Viral Replication • Replication of Animal Viruses – Attachment of animal viruses – Chemical attraction – Animal viruses do not have tails or tail fibers – Have glycoprotein spikes or other attachment molecules that mediate attachment © 2012 Pearson Education Inc. Figure 13.12 Three mechanisms of entry of animal viruses-overview Viral Replication • Replication of Animal Viruses – Synthesis of animal viruses – Requires different strategy depending on its nucleic acid – DNA viruses often enter the nucleus – RNA viruses often replicate in the cytoplasm – Must consider – How mRNA is synthesized – What serves as template for nucleic acid replication © 2012 Pearson Education Inc. Figure 13.13 Synthesis of proteins and genomes in animal RNA viruses-overview Viral Replication • Replication of Animal Viruses – Assembly and release of animal viruses – Most DNA viruses assemble in nucleus – Most RNA viruses develop solely in cytoplasm – Number of viruses produced depends on type of virus and size and initial health of host cell – Enveloped viruses cause persistent infections – Naked viruses are released by exocytosis or lysis © 2012 Pearson Education Inc. Figure 13.14 The process of budding in enveloped viruses Enveloped virion Budding of enveloped virus Viral glycoproteins Viral capsid Cytoplasmic membrane of host Number of infective virions in medium Figure 13.15 Pattern of virion abundance in persistent infections Entry Synthesis and Release of virions assembly Time Attachment Viral Replication ANIMATION Viral Replication: Animal Viruses © 2012 Pearson Education Inc. Viral Replication • Replication of Animal Viruses – Latency of animal viruses – When animal viruses remain dormant in host cells – May be prolonged for years with no viral activity – Some latent viruses do not become incorporated into host chromosome – Incorporation of provirus into host DNA is permanent © 2012 Pearson Education Inc. The Role of Viruses in Cancer • Animal’s genes dictate that some cells can no longer divide or are prevented from unlimited division • Genes for cell division “turned off” or genes inhibiting division “turned on” • Neoplasia – Uncontrolled cell division in multicellular animal – Mass of neoplastic cells is tumor • Benign vs. malignant tumors – Metastasis – Cancers © 2012 Pearson Education Inc. Figure 13.16 The oncogene theory of the induction of cancer in humans Normal state: DNA Protooncogene Represses Gene for repressor mRNA Repressor Result: No cancer First “hit”: Virus inserts promoter DNA Oncogene Represses Gene for repressor mRNA Repressor Result: Still no cancer Second “hit”: Virus inserts into represssor gene DNA Oncogene mRNA Protein No repressor protein because gene is segmented Causes cell division Result: Cancer The Role of Viruses in Cancer • Environmental factors that contribute to the activation of oncogenes – – – – Ultraviolet light Radiation Carcinogens Viruses © 2012 Pearson Education Inc. The Role of Viruses in Cancer • Viruses cause 20–25% of human cancers – Some carry copies of oncogenes as part of their genomes – Some promote oncogenes already present in host – Some interfere with tumor repression – Specific viruses are known to cause ~15% of human cancers – Burkitt’s lymphoma – Hodgkin’s disease – Kaposi’s sarcoma – Cervical cancer © 2012 Pearson Education Inc. Culturing Viruses in the Laboratory • Culturing Viruses in Mature Organisms – In bacteria – In plants and animals • Culturing Viruses in Embryonated Chicken Eggs – Inexpensive, among the largest of cells, free of contaminating microbes, and contain a nourishing yolk • Culturing Viruses in Cell (Tissue) Culture © 2012 Pearson Education Inc. Figure 13.17 Viral plaques in a lawn of bacterial growth on the surface of an agar plate Bacterial lawn Viral plaques Figure 13.18 Inoculation sites for the culture of viruses in embryonated chicken eggs Air sac Injection into chorioallantoic membrane Injection into chorioallantois Injection into embryo Injection into amnion Injection into yolk sac Culturing Viruses in the Laboratory • Culturing Viruses in Cell (Tissue) Culture – Consists of cells isolated from an organism and grown on a medium or in a broth – Two types of cell cultures – Diploid cell cultures – Continuous cell cultures © 2012 Pearson Education Inc. Figure 13.19 An example of cell culture Are Viruses Alive? • Some consider them complex pathogenic chemicals • Others consider them the least complex living entities – Use sophisticated methods to invade cells – Have the ability to take control of their host cell – Are able to replicate themselves © 2012 Pearson Education Inc. Other Parasitic Particles: Viroids and Prions • Characteristics of Viroids – Extremely small, circular pieces of RNA that are infectious and pathogenic in plants – Similar to RNA viruses, but lack capsid – May appear linear due to H bonding © 2012 Pearson Education Inc. Figure 13.20 The RNA strand of the small potato spindle tuber viriod (PSTV) Genome of bacteriophage T7 PSTV Figure 13.21 One effect of viroids on plants Other Parasitic Particles: Viroids and Prions • Characteristics of Prions – Proteinaceous infectious agents – Cellular PrP protein – Made by all mammals – Normal structure with -helices called cellular PrP – Prion PrP – Disease-causing form with -pleated sheets called prion PrP – Prion PrP changes shape of cellular PrP so it becomes prion PrP © 2012 Pearson Education Inc. Other Parasitic Particles: Viroids and Prions ANIMATION Prions: Overview © 2012 Pearson Education Inc. Figure 13.22 The two stable, three-dimensional forms of prion protein (PrP)-overview Other Parasitic Particles: Viroids and Prions • Characteristics of Prions – Normally, nearby proteins and polysaccharides force PrP into cellular shape – PrP mutations result in formation of prion Pr © 2012 Pearson Education Inc. Other Parasitic Particles: Viroids and Prions ANIMATION Prions: Characteristics © 2012 Pearson Education Inc. Other Parasitic Particles: Viroids and Prions • Characteristics of Prions – Prion diseases – Fatal neurological degeneration, fibril deposits in brain, and loss of brain matter – Large vacuoles form in brain – Characteristic spongy appearance – Spongiform encephalopathies – Prions only destroyed by incineration or autoclaving in 1 N NaOH © 2012 Pearson Education Inc. Figure 13.23 The brain of a sheep with the prion disease called scrapie Vacuole Other Parasitic Particles: Viroids and Prions ANIMATION Prions: Disease © 2012 Pearson Education Inc.