Authors: Thompson, Dorothea K.; Palmer, John R.; Daniels, Charles J.

Source: Molecular Microbiology, Volume 33, Number 5, September 1999 , pp. 1081-1092(12)

Publisher: Wiley-Blackwell

Abstract:

Multiple divergent genes encoding the eukaryal-like TFIIB (TFB) transcription initiation factor have been identified in the archaeon Haloferax volcanii. Expression of one of these TFB-encoding genes, referred to here as tfb2, was induced specifically in response to heat shock at the transcription level. A time course for tfb2 induction demonstrated that mRNA levels increased as much as eightfold after 15 min at 60°C. A transcription fusion of the tfb2 promoter region with a stable RNA reporter gene confirmed the heat responsiveness of the tfb2 core promoter, and immunoblot analysis using antibodies generated against a recombinant His-tagged TFB2 showed that the protein levels of one TFB increased slightly in response to elevated temperatures. An archaeal consensus TATA element (5′-TTTATA-3′) was located 110 bp upstream of the translation start site and appeared to be used for both basal and heat shock-induced expression. The long DNA leader region (79 bp) preceding the predicted AUG translation start codon for TFB2 contained a T-rich sequence element located 22 bp downstream of the transcription start site. Using an in vivo transcription termination assay, we demonstrated that this T-rich element can function as a sequence-dependent transcription terminator, which may serve to downregulate expression of the tfb2 gene under both non-heat shock and heat shock conditions.

Document Type: Research article

DOI: http://dx.doi.org/10.1046/j.1365-2958.1999.01551.x

Affiliations: 1: Department of Microbiology, The Ohio State University, 484 West 12th Avenue, Columbus, OH 43210, USA.

Publication date: 1999-09-01

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