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Chapter 18 Microbial Models: The Genetics of Viruses and Bacteria What is a virus? An infectious particle consisting of nucleic acid enclosed in a protein coat. Virus Characteristics 1. 2. 3. 4. 5. Small size Infectious Reproduction Alcohol Resistance Crystallization Size Large - barely visible with the light microscope. Small - down to 20nm. Comment - Much smaller than cells including bacteria. Infectious Viruses cause many diseases. They can be spread from one organism to another. Reproduction Can not reproduce without a host. Viruses are obligate parasites. Alcohol Resistance Not immediately killed by alcohol like cells are. Reason - resistant to dehydration by osmosis. Crystallization Can form crystals. Cells can't do this. Viral Structure 1. Genome - the genetic information. 2. Capsids and Envelopes the outer covering. Viral Genomes Are used to classify virus types. I - ds DNA II - ss DNA III - ds RNA IV-VI - ss RNA Capsid Protein shell that encloses the viral genome. Made from a large number of protein subunits. Number of kinds of subunits usually small. Often geometric in shape. Envelopes Membranes cloaking the capsids of some viruses. Made from host membrane, but may have virus originated glycoproteins added. General Steps for Viral Replication 1. 2. 3. 4. 5. Cell Entry Genome Replication Capsid Protein formation Self-assembly of offspring Exit from Host Cell Entry Locate host by "Lock-andKey” fit between virus proteins and cell receptor molecules. Infection - Begins when viral genome enters the cell. Genome Replication Host is reprogrammed to copy virus genome. Use host DNA polymerase. Use virus enzymes to copy RNA to DNA. Use host's resources to make copies. Capsid Protein Formation Host's protein synthesis machinery used to make virus proteins. Self-Assembly of Offspring genome + capsid -----> virus Particles usually assemble spontaneous. Exit From Host Host cell may burst or lyse, releasing the virus. May "bud-off" host membrane, forming envelopes around the capsids. Bacterial Viruses Best understood of all viruses. May show two types of virus life cycles: Lytic Cycle Lysogenic Cycle Lytic Cycle Virus reproductive cycle that kills the host cell. Note - the previously described virus life cycle was a Lytic Cycle. Ex: T4 Virus on E. coli Uses Lytic Cycle. Example of an Virulent Virus (a virus that only uses the lytic cycle). Has about 100 genes. Completes cycle in 20-30 minutes. Lysogenic Cycle Virus reproduction that doesn't immediately kill the host cell. Viral DNA is inserted into the host DNA, but not expressed. Movie – Lysogenic Cycle Temperate Viruses Viruses with both lytic and lysogenic cycles. l phage - Temperate virus on E. coli. (l = Lambda ) Prophage Virus DNA inserted into the host's DNA. Viral DNA is reproduced with host DNA. "Timebomb" for lytic cycle. Lytic Mode Triggers Switches the host from Lysogenic to Lytic Cycles ex: radiation, chemicals, stress Animal Viruses Belong to several viral classes. Specific Interests: 1. Viruses with envelopes 2. RNA viruses Envelopes made with Host Membranes from: 1. Plasma Membrane 2. Nuclear Membrane Herpes Viruses Use nuclear membrane. Viral DNA integrated into Host DNA as a provirus. Shows both lytic and lysogenic life cycles. RNA Viruses Classes III - VI Class VI - Retrovirus use Reverse Transcriptase to make DNA from an RNA template. Human Immunodeficiency Virus HIV - causes AIDS. Retrovirus from chimps. Destroys the body’s immune system, allowing other diseases to kill. Movie - HIV Other Viral Diseases Measles Polio Smallpox Influenza Treatments for Viruses Vaccines (preventative). Some Drugs Ex: Ara-A Comment Acyclovin - some treatments are working on the reverse transcriptase. Problem Since viruses work within a host cell, they are very difficult to treat. Best Treatment - prevention. Viruses Can also cause cancer Ex: HTLV-1 Papilloma Viruses Plant Viruses Can be an important agricultural problem. Ex: Tobacco Mosaic Virus Tulip Flower Breakage Viroids Infectious particles of naked RNA. Similar to viruses, but lack a capsid. Ex: Coconut Blight Chrysanthemum Wilt Viroids Sequences are similar to Introns. Connection? Prions Infectious protein particles. Cause “mad-cow” and Creutzfeldt-Jakob disease. Protein folding problem? Prion Action Bacteria Genomes DNA circle (Chromosome). Plasmids. - Small circle of DNA that is independent of the chromosome. Carries a small number of traits. E. coli Mutations Binary Fission is not always perfect. Mistakes or mutations are made. Rate: 1x10-7 cell divisions. Rare, but impact is high with the short generation time. Importance Mutations are the prime source of new alleles in bacteria. Bacterial Genetic Recombinations 1. Transformation 2. Transduction 3. Conjugation 4. Plasmids 5. Transposons Be able to discuss a few of these methods. Genetic Recombination Mixes the genetic material into new combinations. Will move new mutations through a population. Transformation Alteration of a cell's DNA by the uptake of foreign DNA. Ex: Griffith's experiment and our lab last semester. Transduction Transfer of genes through phage infections. Conjugation Direct transfer of genetic material between two bacterial cells. Bacterial "sex”. Used to map genetic sequences in bacteria. Plasmids Small circular piece of DNA. Carry many important traits. Ex: Fertility Factor Antibiotic Resistance (R Plasmids) Episome Plasmid that has been incorporated into the bacterial chromosome. Transponsons Transposable genetic elements. Also called "Jumping Genes”. Transposons May change locations within the chromosome, or they may copy into a new location. Don't usually attach to DNA at specific locations, allowing them to scatter genes throughout the genome. Gene Expression in Eukaryotes Important for cellular control and differentiation. Understanding “expression” is a key area in Biology. More details in a future chapter. General Mechanisms 1. Regulate Gene Expression 2. Regulate Enzyme Activity Operon Model Jacob and Monod (1961) Prokaryotic model of gene control. Always on the national AP Biology exam ! Operon Structure 1. Regulatory Gene 2. Operon Area a. Promoter b. Operator c. Structural Genes If Lactose is absent Regulatory Gene Makes Repressor Protein which may bind to the operator. Promoter Attachment sequence on the DNA for RNA polymerase. Operator The "Switch”, binding site for Repressor Protein. If blocked, will not permit RNA polymerase to pass, preventing transcription. Structural Genes Make the enzymes for the metabolic pathway. Lac Operon For digesting Lactose. Inducible Operon - only works (on) when the substrate (lactose) is present. If no Lactose Repressor binds to operator. Operon is "off”, no transcription, no enzymes made If Lactose is absent If Lactose is present Repressor binds to Lactose instead of operator. Operon is "on”, transcription occurs, enzymes are made. If Lactose is present Enzymes Digest Lactose. When enough Lactose is digested, the Repressor can bind to the operator and switch the Operon "off”. Net Result The cell only makes the Lactose digestive enzymes when the substrate is present, saving time and energy. trp Operon Makes Tryptophan. Repressible Operon. If no Tryptophan Repressor protein is inactive, Operon "on” Tryptophan made. “Normal” state for the cell. Tryptophan absent If Tryptophan present Repressor protein is active, Operon "off”, no transcription, no enzymes Result - no Tryptophan made If Tryptophan present Repressible Operons Are examples of Feedback Inhibition. Result - keeps the substrate at a constant level. CAP - positive gene regulation Catabolite Activator Protein Accelerates the level of transcription by working with the RNA polymerase. Uses cAMP as a secondary cell signal. CAP - Mechanism Binds to cAMP. Complex binds to the Promoter, helping RNA polymerase with transcription. Result If the amount of glucose is low (as shown by cAMP) and lactose is present, the lac operon can kick into high gear. Summary Know the general structure of a virus and how a virus reproduces. Know the components of bacteria genomes and some methods for genetic recombination. Summary Know Operon theory ! Know the difference between inducible and repressible enzymes.