In vitro phosphorylation reaction using extracts prepared from cells in the exponential phase of growth and aerial spores of Streptomyces coelicolor displayed the presence of multiply phosphorylated proteins. Effect of protein kinase inhibitors (PKIs) (geldanamycin, wortmannin, apigenin, genistein, roscovitine, methyl 2,5-dihydroxycinnamate, rapamycin, staurosporine) was determined on protein phosphorylation and on germination of spores. The in vitro experiments showed differences in phosphoprotein pattern due to the presence of PKIs. Cultivation of aerial spores with PKIs led to a significant delay in germ tube emergence and filament formation. However, none of the tested PKIs completely blocked the germination process. These results indicate that protein kinases of spores form complex networks sharing common modulating site that plays an important role in proper timing of early developmental events.

• MeSH
• Bacterial Proteins metabolism   MeSH
• Phosphorylation drug effects   MeSH
• Protein Kinase Inhibitors pharmacology   MeSH
• Protein Kinases drug effects   metabolism   MeSH
• Spores, Bacterial drug effects   metabolism   MeSH
• Streptomyces coelicolor drug effects   enzymology   growth & development   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Bacterial Proteins MeSH
• Protein Kinase Inhibitors MeSH
• Protein Kinases 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