Lecturer: IK: Ivo Konopásek, RF: Radovan Fiser, GM: Gabriela Mikusova, PL: Petra Liskova

1. Regulation in bacteria (IK) Regulation of enzyme synthesis, regulation of gene expression in bacteria and archea Regulation of enzyme activity

2. Global Regulation (IK) Global regulation: the answer to environmental stress Stringent response and carbon source limitations Limitations of nitrogen source Phosphate limitation Regulation of transition between aerobic and anaerobic metabolism

3. Environmental stress and taxis (IK) RpoS and global stress response Sigma factors and regulation of stress response Thermal shock Cold shock Oxidative stress Quorum sensing Osmotic shock Chemotaxis

4. Bacterial protein toxins (RF) Toxins acting as superantigens superantigens mechanism of action (binding) examples + producers: SEA, SEB, SECn, SEED, SEE, SEG, SEH (Staphylococcus aureus) SPEA SPEC (Streptococcus pyogenes) SPEB (streptococcal pyrogenic exotoxin B) SMEZ (p. Mitogenic exotoxin Z) MAM (Mycoplasma arthritidis T-cell mitogen) Cell membrane damaging toxins (PFT) General mechanism of action Common features of PFT Examples: RTX toxins (G + / G- bacteria) CyaA - Bordetella pertussis HlyA- (Escherichia coli) "small" PFT (G + bacteria) alpha toxin (Staphylococcus aureus) "large" PFT (G + bacteria) Common features Specific membrane interaction Changes in monomer conformation Oligomer formation examples + producers: PFO - perfringolysin O (Cl. Perfringens) SLO - Streptolysin O (Str. Pyogenes) PLY - pneumolysin (Str. Pneumonia) LLO - Listeriolysin O (L. monocytogenes) Approaches to study pore-forming toxins (PFT) In vitro Model system of black lipid membranes (electrophysiology) Study of PFT using liposomes (fluorescence) Droplet and hydrogel system (electrophysiology + fluorescence) In vivo - Monitoring of Ca2 + (fluorescence)

5. Life of bacteria in biofilm (PL) Basic characteristics and occurrence of biofilm Physiology of cells in biofilm Extracellular matrix Development and dispersion of biofilm Signals affecting biofilm formation Multi-species biofilm

6. Lipids in the membrane of bacteria (GM) Types of bacterial lipids (polar heads, fatty acids, primary structure) Membrane models Non-annular lipids Annular lipids - effect on the function of membrane proteins Function of acidic phospholipids in the membrane Function of phosphatidylethanolamine in membrane Membrane lipids as a target of antibiotics

7. Bacillus subtilis - differentiation and multicellular (GM) Characteristics of Bacillus subtilis Differentiation - cell types and cell fate determination Differentiation triggering signals Regulation of gene expression heterogeneity

8. Non-ribosomal peptide synthesis (GM) Characteristics of NRPS peptides and principle of synthesis Basic domains of synthetase Domain Tailoring Modification of NRPS

9. General mechanisms of bacterial pathogenesis (OC)

10. Heterogeneity of bacterial population in pathogenesis (OC)