抗生素ppt课件（英文精品） molecular mechanisms of microbial pathogenesis and virulence

Molecular Mechanisms of Microbial Pathogenesis and Virulence,Controlling Infectious Diseases The pre-antibiotic Era- Improving Sanitation & hygiene education Typhoid (Salmonella), Plague (Yersina pestis) Syphilis (Treponema pallidum) 2. The antibiotic Era (1940-70)- Defeat of infectious bacteria? 3. Post-antibiotic Era - Emergence and reemergence of infectious diseases. Deaths due to TB (3 million) & Diarrhea (3-4 million) Legionnaires disease (Legionella pneumoniae) Lyme Disease (Borriella burgdoferi) Gastric ulcers & cancer: Helicobacter pyori Campylobacter jejuni 7 strains of E. coli Viral agents:SARS, Morbilli, HIV, Hunta,We have lost control over infectious diseases? Why? Role of Antibiotics: Overuse of antibiotics / antimicrobials has increased drug resistant microbes; no new antibiotics have been discovered in the past 20 yr Some examples of antimicrobial ressistance: Pseudomonas aeruginosa dominate hospital burn units Emergence of multiple (methicillin) resistant staphylococcus aureus (MRSA); Some strains are now resistant to vancomycin, the only effective remaining antibiotic,Decade,2. Improved standard of living: Air conditioners and Legionella 3. Removing and / or substituting environments: SARS, HIV, Lyme disease 4. Aging population, compromised immunity,Microbes adapt to the stress and pressure placed on them 1. Physiological Adaptation Quorum sensing 2. Genetic Adaptation,Various stages of bacteria eucaryote cell interaction during infection,Trojan Horse: quorum sensing,Physiology,Acquiring new genes - Genetic,Quorum sensing - bacterial signaling: Low molecular weight compounds Inhibit macrophage pathogenesis genes not switched on. Legionella in cooling towers,Pathogenic microbes subvert functions of hosts in many ways:Host signaling, host-parasite signaling; parasite signaling,Acquiring New Genes- Horizontal Gene Transfer & Genetic Recombination,1. Horizontal Gene Transfer Mechanisms: Transformation Transduction Conjugation 2. Fate of the Transferred Genetic Material: Incorporated into the hosts genome - the mechanism Not incorporated into the hosts genome 3. Consequences for the Host Cell: New functions,Genetic Recombination is: An exchange between two homologous DNA molecules - general / homologous recombination. What is the mechanism Results in a new combination of genes Mechanism of recombination - E.coli model: Complex, 25 genes: recA genes essential & universal Homologous DNA pair up Donor DNA nicked (nuclease) & displaced (helicase) eg E.coli: RecBCD - nuclease +helicase Reunion of paired segments by strand invasion (Single Strand Binding protein + RecA complex) Exchange of homologous DNA - DNA polymerase & DNA ligase,Recombination results in a new combination of genes Natural process Occurs extensively in all cell types (domains Bacteria, Archaea directly proportional to the distance,Evidence for Horizontal Gene Transfer,Mitochondria - Evidence for lateral gene transfer,Mitochondria Classic evidence for lateral gene transfer and genome miniaturization? Mitochondria are symbiotic bacterium in a eucaryote: multiply by cell division, posses 300-400 proteins in a complex membrane (IM imported into the mitochondria as polypeptides (remember self assembly of cell membranes),Mitochondria genesis and function is a product of both the mitochondria and nuclearencoded genes. Consequently, a mechanism for the coordination of transcription must exist. Is this under mitochondrial (prokaryotic) control,The Human Mitochondria genome database at the URL /,Parasitism also leads to genome miniaturization Host provides metabolic and physiological requirements Mycoplasma genitallium: 580 kb, 470 predicted ORF; Similar to the number of genes which encode human mitochondria. No M. genitallium nuclear encoded genes found in the eucaryotic nucleus. There is a price to pay for the reduced genome and maintaining parasitism: gene addition. Significant number of unique genes code for adhesins, attachment organelles and a number of different membrane-surface antigens directed towards evasion of the immune system,M. peumoniae genome is 816kb (an additional 209 ORFs). How did this difference occur in the same genus? http:/www.zmbh.uni-heidelberg.de/M_pneumoniae/genome/G_Comparison.html Gene transfer, of course!,Microbial Gene Transfer: How are genes transferred? What are the consequences?,Gene transfer in the microbial world Transformation Transductuction Conjugation,Chromosome Plasmid,Involves a donor & a recipient cell DNA is transferred by,Homologous recombination Heritable traits,Transformation,Remember that transformation may have different meanings Cell Biologists interpretation: Transformed cell lines means immortalized cells. This can occur by DNA mutation or by gene transfer mechanisms Microbiologists interpretation: Transformation is one the 3 methods of gene transfer,Dont learn transformation in isolation. Make connects with the knowledge you already have on cell membranes,Transformation: Cell death & DNA fragments released into the environment DNA uptake by related recipient species DNA integration with recipients chromosome Recipient & its progeny cells = genotypically distinct State of the cell, state of DNA, integration of the DNA Competence, DNA uptake, Integration of DNA-Transfection, Artficially induced competence, DNA transfer by electroporation,Recipients should be competent- only some strains naturally competent; artificially induced competency (E.coli) Genetic (regulatory) basis for competency: Membrane-associated DNA binding protein, autolysin, nucleases Bacillus subtilis- Quorum sensing cell number dependent competency Gram -ve: uptake of ds DNA but periplasmic space nucleases renders it ssDNA Gram +ve: dsDNA degraded to ss DNA & then taken up (S. pneumoniae- 10 molecules of 15-20kb dsDNA bind to DNA binding proteins but only 8kb ss DNA enters the cytoplasm) Initial DNA binding is reversible but later becomes irreversible After uptake, the ss DNA binds to competency-specific proteins- nuclease attack proof,Transformation under natural conditions: E. coli is not naturally competent. Uptake artificially induced by electric field or chemicals. ds linear or ds plasmid DNA. Maximum of 20% population transformed. Most human / animal microbes not naturally competent (exception S. pneumoniae 5% of cultivable soil microbes are competent. Possible reason- soil is a harsh extreme environment lacking nutrients. DNA of lysed cells binds to soil, is protected from nucleases and becomes food. In the intestinal tract, DNA is rapidly degraded by nucleases and not available. “Adaptive induced strategy”,Transduction,Transduction: Bacteriophage (phage) mediated gene transfer Bacterium (host) phage specific interaction: phage attaches to host cells surface receptors & injects DNA leaving the capsid outside. Inside the cell, DNA can either: (a) reproduce to form phage and lyse the host to release the phage progeny (process called lytic) OR (b) integrate into the host genome (process called lysogeny). Lysogenic phage may contain phage-encoded toxin genes (botulinum,diptheria, cholera and E. coli O157:H7 bloody diarrhea & kidney failure). Lysogenic phage can be induced into a lytic cycle Some phage progeny released from the lytic may contain host DNA (transducing phage) which is transfered into a new host in the next infection cycle. The foreign host DNA can integrate by homologous recombination or be lost the process is called generalised transduction (see next figure),Generalised transduction,Conjugation,Conjugation Cell to cell contact between a donor and a recipient cell in which genes are transferred. Conjugation process requires different proteins- one protein forms a bridge between the two cells (pili) and another is required for transfer of the gene. The genes are located on plasmids.,Conjugation Understanding Plasmids 1. Physical nature of plasmids 2. Replication of plasmids 3. Cell to cell transfer of plasmids by conjugation 4. Types of plasmids & their biological significance 5. Resistance Plasmids 6. Toxins & other virulence characteristics 7. Bacteriocins,Understanding Plasmids Plasmids are genetic elements replicate independently Plasmids do not have an extracellular form like a virus Thousands of types of plasmids are known; E. coli has over 300 different types 1. Physical nature of plasmids DS DNA Circular (but linear forms are also known) 1 - 1000 kb in size Supercoiled Plasmids can be eliminated - cured; F plasmid by acridine orange Episomes - plasmids that integrate into the host chromosome Isolation and purification possible,Understanding Plasmids 2. Replication of plasmids Involves normal cell enzymes Synchronised event during host cell division 1 to 100 copies per cell- controlled by genes on the plasmid Many different types of plasmids can reside in one cell- regulated by genes on the plasmid eg Borriella burgdoferi- 17 types of circular & linear plasmids. Incompatible (Inc) group plasmids unable to coexist as the replication mechanisms are in common Similar replication mechanism to that for chromosomes - initiated at the origin of replication, unidirectional or bi-directional Small size of plasmids means rapid replication- one-tenth time of cell division Phage phiX174 replicates by rolling circle which has a ss intermediate- ss DNA plasmids Linear plasmids replicate by a priming protein bound to the 5 end,Understanding Plasmids 3. Cell to cell transfer of plasmids by conjugation Conjugation plasmids encode (i) mating pair functions and (ii) DNA transfer (transmissibility) and replication. Two types of types of plasmids are known: (i) self transmissible and (ii) conjugative transposons Self-transmissible Plasmids: Large 30kb plasmids.Can transfer themselves (F+); mobilisable plasmids: Unable to transfer themselves as they lack the ability to form a mating bridge (F-),Conjugative Transposons: Integrate into host chromosome. It can mobilise the transfer of the chromosomal DNA from one cell to another. Strains that transfer high large amounts of chromosomal DNA during conjugation are called high frequency recombinants (Hfr),Origin of transfer,Understanding Plasmids 4. Types of plasmids & their biological significance Remember- essential host functions not part of plasmid genes Pseudomo