'Viruses are obligate intracellular parasites: they reproduce only within a host cell. A virus can only inIect a limited range / type oI host cell 1. Viruses contain no metabolic enzymes or "machinery" Ior protein synthesis.
'Viruses are obligate intracellular parasites: they reproduce only within a host cell. A virus can only inIect a limited range / type oI host cell 1. Viruses contain no metabolic enzymes or "machinery" Ior protein synthesis.
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'Viruses are obligate intracellular parasites: they reproduce only within a host cell. A virus can only inIect a limited range / type oI host cell 1. Viruses contain no metabolic enzymes or "machinery" Ior protein synthesis.
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Chapter 18: Microbial Models: The Genetics oI Viruses and Bacteria
I. What is a virus? Figures 18.1 and 18.2 A. InIectious particles consisting oI nucleic acid (genome) enclosed in a protein coat (capsid), and in some cases, a membranous envelope B. Very small in size C. Some types inIect bacteria (bacteriophage) 1. T series; especially T even phages (T2, T4, and T6) D. Some types inIect plant cells 1. Tobacco mosaic virus (TMV) was Iirst virus discovered E. Animal viruses 1. Adenoviruses- cause colds (as do rhinoviruses) 2. InIluenza viruses- cause inIluenza (aka the Ilu) 3. Human immunodeIiciency virus (HIV)- causes AIDS F. Isolated virusvirus particlevirion II. 'iral genomes (set of genes) A. Unusual in that genome can be double-stranded DNA, single-stranded DNA, double-stranded RNA, or single-stranded RNA B. DNA genomeDNA viruses C. RNA genomeRNA viruses D. Usually single linear or circular molecule oI nucleic acid III.apsids A. Protein shell that encloses viral genome B. DiIIerent shapes C. Many are easily crystallized IV.'iral envelopes A. Found in many animal viruses B. Membranes surrounding capsid C. Derived Irom host cell, but contain additional virally encoded proteins V. 'iral reproduction: Figure 18.3 A. Viruses are obligate intracellular parasites: they reproduce only within a host cell 1. Viruses contain no metabolic enzymes or "machinery" Ior protein synthesis B. A virus can only inIect a limited range/type oI host cell 1. Some viruses can inIect several species, such as the rabies virus 2. Some viruses can only inIect one species C. Animal viruses are usually tissue or cell-type speciIic; that is they will only inIect one particular type oI cell D. Steps oI viral liIe cycle 1. Virus binds to host cell 2. Lock and key Iit between viral protein and host cell surIace receptor 3. Viral genome, via variety oI mechanisms, enters cell 4. Viral genome "commandeers" its host, using host cells machinery to copy the viral genome and synthesize viral proteins E. DNA viruses usually use host cell's DNA polymerases F. RNA viruses must use special virally encoded polymerases (such as reverse transcriptase) VI.acteriophages A. Some double stranded DNA viruses can reproduce by two alternative means B. tic ccle Figure 18.4 1. Phage inIects cell, reproduces, and is released by lysis oI host cell 2. Lytic viruses only reproduce by lytic cycle 3. T4 is a lytic virus C. sogenic ccle Figure 18.5 1. Replicates viral genome without killing cell 2. Viruses with lysogenic cycles called temperate viruses 3. phage 4. In lysogenic cycle, viral genome is in incorporated into host cell's genome by crossing over at a speciIic site . Viral genome is then called a prophage 6. As host cell replicates, viral genome is replicated . At some point, viral genome is activated and initiates lytic cycle VII. nimal viruses A. Very diverse, many modes oI reproduction %able 18.1 B. Viral envelopes Figure 18.6 1. Outer membrane Iound on many animal viruses 2. Helps virus enter the host cell 3. Typically a lipid bilayer (derived Irom host cell plasma membrane) with virally encoded proteins protruding Irom it 4. Virally encoded proteins typically important Ior binding and entering host cell C. erpesvirus envelope derived Irom nuclear envelope 1. Herpesvirus genome integrated into host cell DNA (as provirus) 2. Virus usually remains latent 3. Stress causes virus to become active, provirus leaves genome and intiates viral production 4. Blisters Iorm as a result VIII. # viruses A. Some phages, most plant viruses, and many animals viruses have RNA as genetic material B. RNA is used as genetic material in many diIIerent ways 1. Functions directly as mRNA 2. Functions as template Ior mRNA 3. Functions as template Ior DNA copy oI genome (retroviruses) 4. In retroviruses, inIormation Ilow is: RNA DNA RNA protein a. Reverse transcriptase uses RNA as template Ior DNA synthesis b. HIV liIe cycle Figure 18.7 IX.'iroids and prions A. Viroids are small molecules oI naked RNA that inIect plants B. Prions are inIectious proteins 1. Cause a number oI degenerative brain diseases 2. Mad cow disease 3. Scrapie in sheep 4. CreutzIeldt-Jakob disease in humans . Maybe a misIolded version oI a normal protein, which redirects protein Iolding in inIected cells, generating more misIolded proteins which can inIect other cells Figure 18.9 X. acteria A. Escherichia coli (aka E. coli) are the "laboratory mice" oI molecular biology B. acteria are ver adaptable to their environments 1. Bacteria divide by binary Iusion, preceded by replication oI bacterial chromosome Fig. 18.10 2. Bacteria proliIerate rapidly (20 minutes Ior E. coli) 3. Cannot increase genetic diversity by meiosis and Iertilization 4. Low mutation rate, but mutations can have a signiIicant eIIect on genetic diversity because oI rapid rate oI proliIeration oI E. coli . Genetic recombination also adds genetic diversity to a population C. enetic recombination can produce new bacterial strains 1. Genetic recombination Ior bacteria combining DNA Irom two individuals into a single individual 2. Example oI argtrp- and arg-trp strains interacting to produce argtrp strain Fig. 18.11 3. Genetic recombination can occur by a. %ransformation i. Alteration oI a bacteria's genotype by the uptake oI a naked, Ioreign piece oI DNA ii. Foreign DNA can be incorporated into bacteria's chromosome by crossing over at homologous (similar) sequences iii. II Ioreign DNA is plasmid DNA, does not incorporate into bacterial chromosome b. %ransduction i. Phages carry DNA Irom one bacteria to another ii. eneralized transduction: DNA Irom inIected cell is accidentally packaged into a phage, and transIerred to another bacteria by inIection; O Occurs in a lytic cycle Fig. 18.12 iii. $pecialized transduction: when prophage (oI lysogenic cycle) is excised Irom bacterial genome, it takes a small amount oI the bacterial DNA with it. This DNA is then packaged into a phage and transIerred to another bacteria by inIection c. onjugation i. Conjugation is the direct transIer oI DNA between bacterial cells that are temporarily joined ii. "Maleness" is conIerred by a piece oI DNA called an F Iactor O F Iactor can exist in bacterial chromosome or as a plasmid O F Iactor encodes genes required Ior Iormation oI sex pili O bacteria with F Iactor are F iii. "Female" cells are F- iv. "Male" bacteria Iorm sex pili which attaches to "Iemale" bacteria Fig. 18.13 v. Copy oI F Iactor is transIerred across pili Fig. 18.14 O iI plasmid F Iactor, plasmid is transIerred O iI F Iactor is integrated into the bacterial chromosome, bacterial DNA is also transIerred vi. II F Iactor integrates into bacterial chromosome, bacteria is said to be an HIr cell (Ior high Irequency oI recombination) XI.obile genetic elements A. %ransposon is a piece oI DNA that can move Irom one point to another in bacterial cell's genome B. Transposons do not exist as independent genetic elements (like plasmids) C. Can move DNA within bacterial chromosome or Irom one plasmid to another (i.e. to give multiple drug resistance) D. Some transposons "jump" Irom one location to another; cut and paste transposition E. In replicative transposition, the transposon is copied, and the copy inserts in the new location F. Insertion sequences Fig. 18.15 1. Simplest transposons 2. Contain one gene, transposase, which catalyzes transposition 3. Ends are inverted repeats oI 20-40 base pairs G. Mechanism shown in Fig. 18.16 1. DNA is cut in staggered Iashion by transposase 2. Insertion sequence inserted (also by transposase) 3. DNA polymerase and ligase Iill in DNA and ligate ends 4. DNA next to insertion sequence thus contains direct repeats H. Composite transposons contain additional genes Fig. 18.17 1. Additional genes are "sandwiched" between two insertion sequences that travel together XII. ontrol of bacterial gene expression A. Two levels oI metabolic control Fig. 18.18 1. Cells can vary the numbers oI speciIic enzyme molecules made 2. Regulate gene expression B. Cells can adjust the activity oI the enzymes already present 1. Occurs quickly, as it does not require transcription C. Tryptophan example: iI cell is growing in presence oI tryptophan, it does not need to synthesize it 1. hen tryptophan is present, it inhibits the Iirst enzyme involved in synthesizing tryptophan 2. The presence oI tryptophan also causes cell to stop making the enzymes needed Ior tryptophan synthesis a. Occurs at the level oI transcription XIII. perons A. In bacteria, the genes Ior a particular pathway/metabolic process are clustered together on the chromosome, in a unit called an operon B. A single promoter serves all the genes oI the operon C. The clustered genes constitute a transcription unit- one long mRNA is made 1. This long mRNA is translated into separate polypeptides because the mRNA contains separate start and stop codons Ior each D. An operon has a single on-oII switch that controls the expression oI all the genes 1. This switch is termed an operator 2. Operator is located within the promoter or between the promoter and the genes 3. Controls access oI RNA polymerase to the genes E. This cluster oI genes, promoter and operator is termed an operon F. The operon can be switched oII by a protein called a repressor 1. Repressor is the product oI a regulatory gene, which is not part oI the operon and has its own promoter G. Regulatory genes are transcribed continuously at a low rate H. Many repressors are allosteric molecules, with two shapes: active and inactive I. orepressor is a small molecule that interacts with a repressor to switch an operon oII XIV. %he trp operon A. trp operon is an example oI a repressible operon Figure 18.19 B. trp operon is normall transcribed C. hen tryptophan is present, it binds with the trp repressor, triggering an allosteric change D. The trp repressor with bound tryptophan binds to the operator, shutting oII transcription oI the trp operon E. %rptophan is a corepressor XV. #epressible versus inducible operons A. trp operon is repressible because transcription is inhibited b a specific small molecule (i.e. trptophan) interacts with a regulator protein B. nducible operons are stimulated when a specific small molecule interacts with a regulator protein C. lac operon is an example oI an inducible operon Figure 18.20 D. lac operon encodes enzymes needed Ior metabolism oI lactose (milk sugar) E. lacI is the regulatory gene; it encodes an allosteric repressor that binds to the operator in the absence oI lactose F. hen lactose is present, an isomer, allolactose, binds to the repressor and causes a conIormational change so that the repressor can no longer bind to the operator 1. The lac operon is then transcribed G. Allolactose is an inducer, as it induces transcription oI the operon 1. Also an example oI negative control, because the operon is turned oII by the repressor XVI. !ositive gene regulation Figure 18.21 A. %ranscription of lac operon requires both that lactose be present and that glucose be in short suppl B. II glucose levels are high, cell does not need to synthesize enzymes to catabolize glucose C. Glucose levels detected by interaction oI an allosteric protein with a small organic molecule 1. cyclic AMP accumulates when glucose is absent 2. c! receptor protein (#!) binds c! and is an activator of transcription D. CRP binding site next to promoter oI lac operon 1. CRP plus cAMP bind to this site, making it easier Ior RNA polymerase to bind to the promoter and start transcription