This review summarizes the main results obtained in the fields of general and molecular microbiology and microbial genetics at the Institute of Microbiology of the Academy of Sciences of the Czech Republic (AS CR) [formerly Czechoslovak Academy of Sciences (CAS)] over more than 50 years. Contribution of the founder of the Institute, academician Ivan Málek, to the introduction of these topics into the scientific program of the Institute of Microbiology and to further development of these studies is also included.

• MeSH
• Academies and Institutes history   MeSH
• History, 20th Century MeSH
• Genetics, Microbial history   MeSH
• Molecular Biology history   MeSH
• Check Tag
• History, 20th Century MeSH
• Publication type
• Journal Article MeSH
• Historical Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Geographicals
• Czech Republic MeSH

In vitro phosphorylation of EF-Tu was shown in cell-free extract from dormant spores of Streptomyces coelicolor by a protein kinase present in spores. EF-Tu phosphorylation was observed on both intrinsic S. coelicolor factor and externally added purified EF-Tu from S. aureofaciens, on two isoforms. Putative serine and threonine residues as potential phosphorylation targets were determined in primary sequence and demonstrated on 3D structure model of EF-Tu.

• MeSH
• Electrophoresis, Gel, Two-Dimensional MeSH
• Peptide Elongation Factor Tu chemistry   isolation & purification   metabolism   MeSH
• Phosphorylation MeSH
• Models, Molecular MeSH
• Molecular Sequence Data MeSH
• Protein Kinases metabolism   MeSH
• Amino Acid Sequence MeSH
• Sequence Alignment MeSH
• Spores, Bacterial enzymology   metabolism   MeSH
• Streptomyces aureofaciens metabolism   MeSH
• Streptomyces coelicolor metabolism   MeSH
• Protein Structure, Tertiary MeSH
• Imaging, Three-Dimensional MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Peptide Elongation Factor Tu MeSH
• Protein Kinases MeSH

We cloned EF-Tu from Streptomyces aureofaciens on a pET plasmid and overproduced it using the T7 RNA polymerase system in Escherichia coli. Streptomyces EF-Tu represented more than 40% of the total cell protein and was stored mostly in inclusion bodies formed apically at both ends of E. coli cells. Analysis of the inclusion bodies by transmission and scanning electron microscopy did not reveal any internal or surface ultrastructures. We developed the method for purification of S. aureofaciens EF-Tu from isolated inclusion bodies based on the ability of the protein to aggregate spontaneously. EF-Tu present in inclusion bodies was not active in GDP binding. Purified protein showed a similar charge heterogeneity as EF-Tu isolated from the mycelium of S. aureofaciens and all of the isoforms reacted with EF-Tu antibodies. All isoforms also reacted with monoclonal antibodies against O-phosphoserine and O-phosphothreonine.

• MeSH
• Electrophoresis, Gel, Two-Dimensional MeSH
• Bacterial Proteins genetics   immunology   isolation & purification   metabolism   MeSH
• Inclusion Bodies ultrastructure   MeSH
• Electrophoresis, Polyacrylamide Gel MeSH
• Peptide Elongation Factor Tu genetics   immunology   isolation & purification   metabolism   MeSH
• Escherichia coli genetics   metabolism   MeSH
• Gene Expression MeSH
• Genetic Vectors MeSH
• Guanosine Diphosphate metabolism   MeSH
• Cloning, Molecular MeSH
• Microscopy, Electron, Scanning MeSH
• Antibodies, Monoclonal immunology   MeSH
• Plasmids MeSH
• Protein Processing, Post-Translational * MeSH
• Protein Isoforms immunology   MeSH
• Recombinant Proteins immunology   isolation & purification   metabolism   MeSH
• Streptomyces aureofaciens genetics   MeSH
• Microscopy, Electron, Transmission MeSH
• Protein Binding MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Bacterial Proteins MeSH
• Peptide Elongation Factor Tu MeSH
• Guanosine Diphosphate MeSH
• Antibodies, Monoclonal MeSH
• Protein Isoforms MeSH
• Recombinant Proteins MeSH

Protein kinases can be classified into two main superfamilies on the basis of their sequence similarity and substrate specificity. The protein His kinase superfamily which autophosphorylate a His residue, and superfamily Ser/Thr and Tyr protein kinases, which phosphorylate Ser, Thr or Tyr residues. During the last years genes encoding Ser/Thr protein kinases have been identified in several microorganisms. Phosphorylation of proteins on Ser/Thr residues can be involved in many functions of prokaryotic cells including cell differentiation, signal transduction and protein biosynthesis. Phosphorylation of prokaryotic protein-synthesizing systems showed that the phosphorylation of initiation and elongation factors is subject to alteration during cell differentiation or bacteriophage infection. Protein kinase associated with ribosomes of streptomycetes phosphorylate the elongation factor Tu and 11 ribosomal proteins even in bacteriophage-uninfected cells. After phosphorylation of ribosomal proteins, ribosomes lose about 30% of their activity at the translation of poly(U).

• MeSH
• Bacterial Proteins metabolism   MeSH
• Peptide Elongation Factor Tu metabolism   MeSH
• Phosphorylation MeSH
• Molecular Sequence Data MeSH
• Protein Kinases metabolism   MeSH
• Ribosomal Proteins metabolism   MeSH
• Ribosomes metabolism   MeSH
• Amino Acid Sequence MeSH
• Streptomyces enzymology   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Names of Substances
• Bacterial Proteins MeSH
• Peptide Elongation Factor Tu MeSH
• Protein Kinases MeSH
• Ribosomal Proteins MeSH

Prokaryotic cells contain proteins which form extended chains or multimers that oscillate between monomers and oligomers of varying length. Hydrolysis of nucleoside triphosphates combined with site-specific disposition of substrates and products to monomers and multimers is the driving force of dynamic instability of these molecules. Polymeric structures are connected in some manner to a variety of signaling systems that adhere to the polymeric matrix, including the GTP-binding protein(s), protein kinases and phosphatases, and other proteins or systems that communicate between the cytoplasmic membrane and the cytosol. Flexible organization allowing regulated dynamic movement is one of the key elements in all living cells. In eukaryotic cells actin and tubulin are the two main components of dynamically controlled spatial system. These proteins are noteworthy for their ability to polymerize, reversibly, into filaments or microtubules in association with hydrolysis of ATP or GTP, respectively. As such, they regulate most of the mechanics of cell movement including cell division, cell differentiation, phagocytosis and other dynamic phenomena. Recent evidence revealed that microbial cells create functional domains at specific sites of the cells and can form cytoplasmic tubules and fibers.

• MeSH
• Bacteria growth & development   metabolism   MeSH
• Bacterial Proteins metabolism   MeSH
• Cytoskeletal Proteins * MeSH
• Phosphorylation MeSH
• Bacterial Physiological Phenomena * MeSH
• Microtubules metabolism   MeSH
• Movement MeSH
• Polymers metabolism   MeSH
• GTP-Binding Proteins chemistry   metabolism   MeSH
• Tubulin chemistry   metabolism   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Names of Substances
• Bacterial Proteins MeSH
• Cytoskeletal Proteins * MeSH
• FtsZ protein, Bacteria MeSH Browser
• Polymers MeSH
• GTP-Binding Proteins MeSH
• Tubulin MeSH