Genetic Basis of Variation in Bacteria
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1 Mechanisms of Infectious Disease Fall 2009 Genetics I Jonathan Dworkin, PhD Department of Microbiology jonathan.dworkin@columbia.edu Genetic Basis of Variation in Bacteria I. Organization of genetic material in bacteria a. chromosomes b. plasmids II. Genetic variation: Source a. point mutation b. DNA rearrangements III. Genetic variation: Transmission a. transformation b. transduction c. conjugation IV. Genetic variation: Implications for pathogenesis
2 Genetic Basis of Variation in Bacteria I. Organization of genetic material in bacteria a. chromosomes b. plasmids II. Genetic variation: Source a. point mutation b. DNA rearrangements III. Genetic variation: Transmission a. transformation b. transduction c. conjugation IV. Genetic variation: Implications for pathogenesis Genetic Basis of Variation in Bacteria I. Organization of genetic material in bacteria a. chromosomes b. plasmids II. Genetic variation: Source a. point mutations b. DNA rearrangements III. Genetic variation: Transmission a. transformation b. transduction c. conjugation IV. Genetic variation: Implications for pathogenesis
3 Genetic Basis of Variation in Bacteria I. Organization of genetic material in bacteria a. chromosomes b. plasmids II. Genetic variation: Source a. point mutations b. DNA rearrangements III. Genetic variation: Transmission a. transformation b. transduction c. conjugation IV. Genetic variation: Implications for pathogenesis Genetic Basis of Variation in Bacteria I. Organization of genetic material in bacteria a. chromosomes b. plasmids II. Genetic variation: Source a. point mutations b. DNA rearrangements III. Genetic variation: Transmission a. transformation b. transduction c. conjugation IV. Genetic variation: Implications for pathogenesis
4 Genetic basis of variation: Griffiths (1928) Genetic basis of variation: Griffiths (1928)
5 Genetic basis of variation: Avery et al. (1944) Genetic basis of variation: Hershey and Chase (1952)
6 Genetic Basis of Variation in Bacteria I. Organization of genetic material in bacteria a. chromosomes b. plasmids II. Genetic variation: Source a. point mutation b. DNA rearrangements III. Genetic variation: Transmission a. transformation b. transduction c. conjugation IV. Genetic variation: Implications for pathogenesis Organization of genetic material in bacteria: chromosomes Most bacteria contain a single chromosome (+ extrachromosomal elements) Some bacteria have been found also to contain 2-3 replicons which can be considered either megaplasmids or minichromosomes e.g. 3.0 Mb and 0.9 Mb replicons in Rhodobacter sphaeroides A few bacterial genera contain >1 chromosome e.g. 2.1 Mb and 1.2 Mb chromosomes in Brucella Some bacteria harbour large replicons essential for survival in a specific ecological niche but not under laboratory conditions e.g. 1.4 Mb and 1.7 Mb replicons in Rhizobium meliloti are required for plant symbiosis
7 Organization of genetic material in bacteria: chromosomes Organization of genetic material in bacteria: chromosomes Copyright 2004 by the National Academy of Sciences
8 Organization of genetic material in bacteria: plasmids Extrachromosomal Circular or linear 2 kb to hundreds of kb in size Non-essential May carry supplemental genetic information or may be cryptic Employ host functions for most of DNA metabolism Chromosome Plasmid Organization of genetic material in bacteria: plasmids Virulence Replication Maintenance Transfer Antibioticresistance
9 Organization of genetic material in bacteria: plasmids Transposon genes Replication and maintenance Antibiotic resistance Other Organization of genetic material in bacteria: plasmids Examples of naturally-occuring plasmids and relevant features Plasmid Host Plasmid size (kb) Relevant feature pt181 Staphylococcus aureus 4.4 Tetracycline resistance ColE1 Escherichia coli 6.6 Colicin production and immunity pgkl2 Kluyveromyces lactis b 13.5 Killer plasmid pamβ1 Enterococcus faecalis 26.0 Erythromycin resistance psk41 Staphylococcus aureus 46.4 Multidrug resistance pbm4000 Bacillus megaterium 53.0 rrna operon pi258 Staphylococcus aureus 28.0 Metal ion resistance pslt Salmonella enterica subsp. typhimurium 93.9 Virulence determinants pmt1 Yersinia pestis Virulence determinants padp-1 Pseudomonas sp Atrazine (herbicide) catabolism pww0 Pseudomonas putida Aromatic hydrocarbon degradation pbtoxis Bacillus thuringiensis subsp. israelensis Mosquito larval toxicity px01 Bacillus anthracis Exotoxin production psol1 Clostridium acetobutylicum Solvent production psymb Sinorhizobium meliloti Functions associated with plant symbiosis
10 Genetic Basis of Variation in Bacteria I. Organization of genetic material in bacteria a. chromosomes b. plasmids II. Genetic variation: Source a. point mutations b. DNA rearrangements III. Genetic variation: Transmission a. transformation b. transduction c. conjugation IV. Genetic variation: Implications for pathogenesis Sources of genetic variation: point mutations
11 Sources of genetic variation: point mutations Sources of genetic variation: point mutations
12 Sources of genetic variation: DNA rearrangements Insertion sequence (IS) elements: 1. Simplest type of transposable element found in bacterial chromosomes and plasmids. 2. Encode only genes for mobilization and insertion. 3. Range in size from 768 bp to 5 kb. 4. IS1 first identified in E. coli s glactose operon is 768 bp long and is present with 4-19 copies in the E. coli chromosome. 5. Ends of all known IS elements show inverted terminal repeats (ITRs). Sources of genetic variation: DNA rearrangements Integration of IS element in chromosomal DNA.
13 Sources of genetic variation: DNA rearrangements Sources of genetic variation: DNA rearrangements
14 Sources of genetic variation: frequency of occurrence Genetic Basis of Variation in Bacteria I. Organization of genetic material in bacteria a. chromosomes b. plasmids II. Genetic variation: Source a. point mutations b. DNA rearrangements III. Genetic variation: Transmission a. transformation b. transduction c. conjugation IV. Genetic variation: Implications for pathogenesis
15 Transmission of genetic variation Lengths of bars denote the amount of protein-coding DNA. For each bar, the native DNA is blue; foreign DNA identifiable as mobile elements, including transposons and bacteriophages, is yellow, and other foreign DNA is red. The percentage of foreign DNA is noted to the right of each bar. 'A' denotes an Archaeal genome.
16 Transmission of genetic variation: antibiotic resistance V. M. D'Costa et al. Science (2006) Fig. 1. Antibiotic resistance profiling of 480 soil-derived bacterial isolates Transmission of genetic variation: pathogenesis
17 Transmission of genetic variation: Luria-Delbruck test Transmission of genetic variation: Luria-Delbruck test
18 Transmission of genetic variation: Luria-Delbruck test Linear transmission of genetic variation
19 Horizontal transmission of genetic variation Transmission of genetic variation: mechanisms
20 Transmission of genetic variation: transformation Transmission of genetic variation: transformation Gene transfer resulting from the uptake of DNA from a donor. g p Factors affecting transformation DNA size and state Sensitive to nucleases Competence of the recipient (Bacillus, Haemophilus, Neisseria, Streptococcus) Competence factor Induced competence
21 Transmission of genetic variation: transformation Transmission of genetic variation: transformation
22 Transmission of genetic variation: transformation Transmission of genetic variation: mechanisms
23 Genetic Basis of Variation in Bacteria I. Organization of genetic material in bacteria a. chromosomes b. plasmids II. Genetic variation: Source a. point mutation b. DNA rearrangements III. Genetic variation: Transmission a. transformation b. transduction c. conjugation IV. Genetic variation: Implications for pathogenesis Genetic Basis of Variation in Bacteria I. Organization of genetic material in bacteria a. chromosomes b. plasmids II. Genetic variation: Source a. point mutations b. DNA rearrangements III. Genetic variation: Transmission a. transformation b. transduction c. conjugation IV. Genetic variation: Implications for pathogenesis
24 Genetic Basis of Variation in Bacteria I. Organization of genetic material in bacteria a. chromosomes b. plasmids II. Genetic variation: Source a. point mutations b. DNA rearrangements III. Genetic variation: Transmission a. transformation b. transduction c. conjugation IV. Genetic variation: Implications for pathogenesis
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