Structure and expression of elongation factor Tu from Bacillus stearothermophilus
Language English Country Netherlands Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
9769211
DOI
10.1006/jmbi.1998.2102
PII: S0022-2836(98)92102-7
Knihovny.cz E-resources
- MeSH
- Aminoglycosides * MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Drug Resistance, Microbial MeSH
- DNA, Bacterial MeSH
- Peptide Elongation Factor Tu biosynthesis chemistry genetics MeSH
- Geobacillus stearothermophilus drug effects genetics metabolism MeSH
- Protein Conformation MeSH
- Models, Molecular MeSH
- Molecular Sequence Data MeSH
- Pyridones pharmacology MeSH
- Amino Acid Sequence MeSH
- Base Sequence MeSH
- Sequence Alignment MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Aminoglycosides * MeSH
- Anti-Bacterial Agents MeSH
- DNA, Bacterial MeSH
- Peptide Elongation Factor Tu MeSH
- mocimycin MeSH Browser
- pulvomycin MeSH Browser
- Pyridones MeSH
The tuf gene coding for elongation factor Tu (EF-Tu) of Bacillus stearothermophilus was cloned and sequenced. This gene maps in the same context as the tufA gene of Escherichia coli str operon. Northern-blot analysis and primer extension experiments revealed that the transcription of the tuf gene is driven from two promoter regions. One of these is responsible for producing a 4.9-kb transcript containing all the genes of B. stearothermophilus str operon and the other, identified adjacent to the stop codon of the fus gene and designated tufp, for producing a 1.3-kb transcript of the tuf gene only. In contrast to the situation in E. coli, the ratio between the transcription products was found to be about 10:1 in favour of the tuf gene transcript. This high transcription activity from the tufp promoter might be accounted for by the presence of an extremely A+T-rich block consisting of 29 nucleotides which immediately precedes the consensus -35 region of the promoter. A very similar tuf gene transcription strategy and the same tufp promoter organization with the identical A/T block were found in Bacillus subtilis. The tuf gene specifies a protein of 395 amino acid residues with a molecular mass of 43,290 Da, including the N-terminal methionine. A computer-generated three-dimensional homology model shows that all the structural elements essential for binding guanine nucleotides and aminoacyl-tRNA are conserved. The presence of serine at position 376 and a low affinity for kirromycin determined by zone-interference gel electrophoresis (Kd approximately 8 microM) and by polyacrylamide gel electrophoresis under non-denaturing conditions are in agreement with the reported resistance of this EF-Tu to the antibiotic. The replacement of the highly conserved Leu211 by Met was identified as a possible cause of pulvomycin resistance.
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