Expression of proteins and protein kinase activity during germination of aerial spores of Streptomyces granaticolor.
Dormant aerial spores of Streptomyces granaticolor contain pre-existing pool of mRNA and active ribosomes for rapid translation of proteins required for earlier steps of germination. Activated spores were labeled for 30 min with [35S]methionine/cysteine in the presence or absence of rifamycin (400 microg/ml) and resolved by two-dimensional electrophoresis. About 320 proteins were synthesized during the first 30 min of cultivation at the beginning of swelling, before the first DNA replication. Results from nine different experiments performed in the presence of rifamycin revealed 15 protein spots. Transition from dormant spores to swollen spores is not affected by the presence of rifamycin but further development of spores is stopped. To support existence of pre-existing pool of mRNA in spores, cell-free extract of spores (S30 fraction) was used for in vitro protein synthesis. These results indicate that RNA of spores possesses mRNA functionally competent and provides templates for protein synthesis. Cell-free extracts isolated from spores, activated spores, and during spore germination were further examined for in vitro protein phosphorylation. The analyses show that preparation from dormant spores catalyzes phosphorylation of only seven proteins. In the absence of phosphatase inhibitors, several proteins were partially dephosphorylated. The activation of spores leads to a reduction in phosphorylation activity. Results from in vitro phosphorylation reaction indicate that during germination phosphorylation/dephosphorylation of proteins is a complex function of developmental changes.
Mikulík
Karel
K
Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídenská 1083, 4, 142 20, Prague, Czech Republic.
Bobek
Jan
J
Bezousková
Silvia
S
Benada
Oldrich
O
Kofronová
Olga
O
D015180:Electrophoresis, Gel, Two-Dimensional; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D015966:Gene Expression Regulation, Fungal; D010766:Phosphorylation; D014176:Protein Biosynthesis; D011494:Protein Kinases / Q000378:metabolism*; D012331:RNA, Fungal / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012294:Rifamycins / Q000494:pharmacology; D013172:Spores, Fungal / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism* / Q000502:physiology
D005656:Fungal Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D012294:Rifamycins; D011494:Protein Kinases