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Expression of the gap gene encoding glyceraldehyde-3-phosphate dehydrogenase of Streptomyces aureofaciens requires GapR, a member of the AraC/XylS family of transcriptional activators

O. Spruanský¹

B. euchová¹

Institute of Molecular Biology, Slovak Academy of Sciences, Dubravská cesta 21, 842 51 Bratislava, Slovak Republic¹

Author for correspondence: J. Kormanec. Tel: +421 7 5941 2432. Fax: +421 7 5477 2316. e-mail: umbijkor{at}savba.sk

Expression of the gap gene encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is developmentally regulated, and induced by glucose in Streptomyces aureofaciens. A gene, gapR, encoding a protein similar to the AraC/XylS family of bacterial transcriptional regulators was identified upstream of gap. The gapR gene was constitutively expressed from a single promoter during the course of differentiation. By integrative transformation, via double crossover, a stable null mutant of the gapR gene was obtained. The mutation only slightly affected growth, and had no effect on differentiation of S. aureofaciens. However, transcription of the GAPDH-encoding gap gene was substantially reduced in the S. aureofaciens gapR null mutant, irrespective of carbon source used. Though GAPDH activity was about 1·5-fold lower in the mutant, the substantial enzyme activity remained, suggesting the presence of a second GAPDH which is sufficient to ensure growth. The GapR protein, overproduced in Escherichia coli, was shown to bind upstream of the gap-P promoter region. The results indicate a direct role of GapR in regulation of gap expression in S. aureofaciens.

Keywords: glucose induction, differentiation, GAPDH, promoter, S1-nuclease mapping

Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; tsp, transcription start point(s); wt, wild-type

The GenBank/EMBL/DDBJ accession number for the sequence described in this paper is U21191.

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