Sequencing of the tuf1 gene and the phosphorylation pattern of EF-Tu1 during development and differentiation in Streptomyces collinus producing kirromycin.
We have cloned and sequenced the tuf1 gene from a kirromycin-producing strain of Streptomyces collinus. The gene encodes a polypeptide of 396 amino acid residues with a molecular weight of 43,849. The protein shows 97% identity with EF-Tu1 of S. coelicolor and is sensitive to kirromycin. EF-Tu-dependent translation of poly(U) was reduced to 50% in the presence of 0.25 microM kirromycin. Using high resolution two-dimensional electrophoresis and specific immunodetection with monoclonal antibodies we found that the EF-Tu1 is phosphorylated on threonine and that serine is the second phosphate-accepting amino acid. EF-Tu1 phosphorylated on threonine and serine residues was detected among the S150 supernatant proteins of vegetative cells, aerial mycelium and spores. The level of phosphorylated EF-Tu1 varied during the growth and differentiation.
Mikulík
K
K
Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague.
Zhulanova
E
E
D000595:Amino Acid Sequence; D001483:Base Sequence; D004269:DNA, Bacterial / Q000737:chemistry; D004352:Drug Resistance, Microbial; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D007163:Immunosorbent Techniques; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D010444:Peptide Elongation Factor Tu / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism; D010769:Phosphothreonine / Q000378:metabolism; D011728:Pyridones / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
D004269:DNA, Bacterial; D011728:Pyridones; D010769:Phosphothreonine; D010768:Phosphoserine; D010444:Peptide Elongation Factor Tu; C008536:mocimycin