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Chapter 18 The Genetics of Viruses and Bacteria Introduction • Viral/bacterial studies led to understanding mechanisms of heredity due to simplicity (viruses – nucleic acid & protein coat only) Studies led to: • better understanding of disease • emergence of biotechnology Discovery of viruses • 1883 A. Mayer – studying tobacco mosaic disease (thought to be carried by unusually small bacteria unseen through microscope) • 1897 Beijerenck – discovered infectious agent could reproduce, therefore, not just a bacterial toxin • 1935 Stanley – crystallized infectious particle; then others could be identified with the aid of the electron microscope Structure of viruses • very small (20 nm smallest) • virion: nucleic acid & protein coat called a capsid • genome can be ds DNA, ssDNA, dsRNA, ssRNA • linear or circular NA (4 - 700 genes) • acellular Identified by shape • capsid – protein coat may be: 1) rod (helical) ex: TMV 2) polyhedral ex: adenovirus 3) complex (combination) ex: T4 bacteriophage • Some have viral envelopes, membranes cloaking their capsids derived from membrane of host cell ex: HIV Viral Reproduction Lytic cycle (virulent viruses) p.332 1) adsorption (attachment) – virus attaches to host cell 2) entry – viral NA enters host cell 3) replication – virus NA takes over host NA & makes new viral NA & viral protein 4) assembly – viral NA & proteins are joined to make new viruses 5) release – viruses break out & destroy host cell (cell is lysed) Lysogenic cycle (temperate viruses) p. 333 1) adsorption 2) entry – viral NA is integrated into host NA called a prophage or provirus -may remain latent in host cell or may be triggered to complete steps like the lytic cycle Possible triggers may include: • Stress • Increased temperature ex: herpes simplex I virus (oral) Animal viruses • Enveloped viruses may have easier access into cells • Herpes viruses derive the envelope from nuclear membrane Retroviruses have the most complicated life cycles • Contain reverse transcriptase RNA a DNA ex: HIV • In some cases, viral damage is easily repaired (respiratory epithelium after a cold), but in others, infection causes permanent damage (nerve cells after polio) Emergent viruses • HIV • New influenza viruses • Ebola due to : 1) mutation of existing viruses (flu strains) 2) spread of existing viruses from 1 species to another (bird flu, hantavirus) 3) dissemination of a viral disease from a small isolated population (AIDS) • Viruses appear to cause certain human cancers -hepatitis B – liver cancer -Epstein-Barr virus – several cancers including Burkitt’s lymphoma -Papilloma viruses – cervical cancer -HTLV -1 – adult leukemia Plant viruses • serious agricultural pests (stunt growth, diminish yield) • spread easily • most are RNA viruses Origin • taxonomic puzzle; they do evolve, but they are not cells Bacterial genome • circular chromosome with a few associated proteins • accessory genes found on smaller rings of DNA called plasmids • replication is bidirectional from a single origin Reproduction in bacteria • Asexually: binary fission – splitting in two • Sexually: 1) transformation – gene transfer where bacterial cell assimilates foreign DNA from its environment Reproduction in bacteria 2) transduction – gene transfer from one bacterium to another by a phage a) generalized – random pieces of host’s DNA are packaged within phage capsid b) specialized – prophage takes piece of bacterial chromosome with it Reproduction in Bacteria 3) conjugation – direct transfer of genes between 2 temporarily joined bacteria Antibiotic resistance • bacterial genes may code for enzymes that destroy certain antibiotics • these genes are carried by plasmids known as R plasmids (R = resistance) • resistant bacteria survive to pass on these genes (causes increase in bacterial strains that are antibiotic resistant) • R plasmids can be transferred by conjugation episomes – plasmids that can integrate into bacterial chromosome Transposons transposons – described by Barbara McClintock mobile segments of DNA that may move within a chromosome & to & from plasmids a) conservative – changes location without replicating first b) replicative - replicates, remaining in its original position, also inserting in a new location (can move genes to totally new areas) Insertion sequences • simplest transposons • consist only of DNA necessary for transposition (transposase) • IS flanked by inverted repeats (noncoding segments 20-40 nucleotides long) • enzyme (transposase) recognizes these inverted repeats; enzyme binds to catalyze cutting and resealing • While insertion sequences may not benefit bacteria in any specific way, composite transposons may help bacteria adapt to new environments. • For example, repeated movements of resistance genes by composite transposition may concentrate several genes for antibiotic resistance onto a single R plasmid. • In an antibiotic-rich environment, natural selection factors bacterial clones that have built up composite R plasmids through a series of transpositions. • Transposable genetic elements are important components of eukaryotic genomes as well. • In the 1940s and 1950s Barbara McClintock investigated changes in the color of corn kernels. – She postulated that the changes in kernel color only made sense if mobile genetic element moved from other locations in the genome to the genes for kernel color. – When these “controlling elements” inserted next to the genes responsible for kernel color, they would activate or inactivate those genes. – In 1983, more than 30 years after her initial breakthrough, Dr. McClintock received a Nobel Prize for her discovery. • The control of gene expression enables individual bacteria to adjust their metabolism to environmental change • An individual bacterium, locked into the genome that it has inherited, can cope with environmental fluctuations by exerting metabolic control. – First, cells vary the number of specific enzyme molecules by regulating gene expression. – Second, cells adjust the activity of enzymes already present (for example, by feedback inhibition). • For example, the tryptophan biosynthesis pathway demonstrates both levels of control. – If tryptophan levels are high, some of the tryptophan molecules can inhibit the first enzyme in the pathway. – If the abundance of tryptophan continues, the cell can stop synthesizing additional enzymes in this pathway by blocking transcription of the genes for these enzymes. Fig. 18.19 Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings Operator • On – binding of activator protein stimulates transcription (usually the case) • Off – binding of specific repressor protein shuts off transcription constitutive genes – unregulated genes (always needed by cell) Repressible operon • Inhibited by an anabolic end product (such as trp - tryptophan) • trp operon is an example • tryptophan is a corepressor that binds to repressor protein, changing it to active state which switches off trp operon • When trp decreases, repressor protein is no longer bound; operator not repressed, RNA polymerase attaches to promoter & trp synthesis continues Inducible operon • function in catabolic pathways • inducer (substrate for pathway) derepresses operon by inhibiting the otherwise active repressor protein • example is lac operon (lactose) • has 3 structural genes, repressor innately active, binding to lac operator & switching off the operon • allolactose (lactose isomer) acts as inducer, binds to and inactivates repressor protein so operon can be transcribed Repressible vs. Inducible Repressible • • • • • inhibited by anabolic end product inherently “on” innately inactive repressor under negative control ex: trp operon Repressible vs. inducible Inducible • • • • • function in catabolic pathway inherently “off” innately active repressor under negative & positive control ex: lac operon • Both of these are examples of negative control – operons are switched off by the active form of the repressor protein • Positive control – when an activator molecule interacts directly with the genome to switch transcription on • Positive gene control occurs when an activator molecule interacts directly with the genome to switch transcription on. • Even if the lac operon is turned on by the presence of allolactose, the degree of transcription depends on the concentrations of other substrates. – If glucose levels are low (along with overall energy levels), then cyclic AMP (cAMP) binds to cAMP receptor protein (CRP) which activates transcription. • The cellular metabolism is biased toward the utilization of glucose. • If glucose levels are sufficient and cAMP levels are low (lots of ATP), then the CRP protein has an inactive shape and cannot bind upstream of the lac promotor. – The lac operon will be transcribed but at a low level. Fig. 18.22b Copyright © 2002 Pearson Education, Inc., publishing as Benjamin Cummings • For the lac operon, the presence / absence of lactose (allolactose) determines if the operon is on or off. • Overall energy levels in the cell determine the level of transcription, a “volume” control, through CRP. • CRP works on several operons that encode enzymes used in catabolic pathways. – If glucose is present and CRP is inactive, then the synthesis of enzymes that catabolize other compounds is slowed. – If glucose levels are low and CRP is active, then the genes which produce enzymes that catabolize whichever other fuel is present will be transcribed at high levels.