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Nucleic Acids Research, 1995, Vol. 23, No. 10 1737-1743
© 1995

MOLECULAR BIOLOGY

Genetic and molecular analysis of the tRNA-tufB operon of the myxobacterium Stigmatella aurantiaca

Laure Bremaud1, Christophe Fremaux1,2, Soumaya Laalami1 and Yves Cenatiempo1,*

1lnstitut de Biologie Moléculaire et d'lngénierie Génétique—CNRS URA 1172, Université de Poitiers 40 Avenue du Recteur Pineau, 86022 Poitiers Cedex, France 2Texel, groupe Rhône-Poulenc Zl de Buxières, BP 10, 86220 Dangé-St Romain, France

* To whom correspondence should be addressed

Received January 31, 1995. Revised March 30, 1995. Accepted March 30, 1995.

The tufB gene, encoding elongation factor Tu (EF-Tu), from the myxobacterium Stigmatella aurantiaca was cloned and sequenced. It Is preceded by four tRNA genes, the first ever described In myxobacterla. The tRNA synthesized from these genes and the general organization of the locus seem Identical to that of Escherlchla coll, but differences of potential Importance were found In the tRNA sequences and In the Intergenic regions. The primary structure of EF-Tu was deduced from the tufB DNA sequence. The factor is composed of 396 amino acids, with a predicted molecular mass of 43.4 kDa, which was confirmed by expression of tufB in maxicells. Sequence comparisons between S.aurantlaca EF-Tu and other bacterial homologues from E.coli, Salmonella typhlmurlum and Thermus thermophllus displayed extensive homo-logies (75.9%). Among the variable positions, two Cys residues probably involved in the temperature sensitivity of E.coli and S.typhimurium EF-Tu are replaced in T.thermophilus and S.aurantlaca EF-Tu. Since two or even three tuf genes have been described in other bacterial species, the presence of multiple tuf genes was sought for. Southern and Northern analysis are consistent with two tuf genes in the genome of S.aurantlaca. Primer extension experiments indicate that the four tRNA genes and tufB are organized in a single operon.


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