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Microbiology 142 (1996), 2561-2566; DOI  10.1099/00221287-142-9-2561
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An upstream activating sequence containing curved DNA involved in activation of the Clostridium perfringens plc promoter

Chieko Matsushita1, Osamu Matsushita1, Seiichi Katayama1, Junzaburo Minami1, Kenichi Takai2 and Akinobu Okabe1,3

Department of Microbiology, Kagawa Medical School, 1750-1, Miki-cho, Kita-gun, Kagawa 761-07, Japan
Okayama Central Hospital, 2-18-19, Hokan-cho, Okayama 700, Japan

3Author for correspondence: A. Okabe. Tel: +81 878 98 5111. Fax: +81 878 98 7109. e-mail: microbio@kms.ac.jp

ABSTRACT

The plc gene, which encodes phospholipase C ({alpha}-toxin) of Clostridium perfringens, possesses three poly(A) tracts forming an intrinsically curved DNA region immediately upstream of the promoter. The in vivo transcriptional activity of the plasmid-borne plc gene was stimulated by this curved-DNA-containing sequence, depending on its proper linear and rotational orientation. The in vitro transcriptional activity of the plc gene was also stimulated by the upstream sequence. In addition, the stimulatory effect of the sequence and the degree of DNA bending were greater at lower temperature, as was demonstrated by both in vitro and in vivo transcription assays, and a gel-mobility assay, respectively. A similar temperature effect was also observed with the chromosomal plc gene. These observations suggest that the upstream DNA curvature per se stimulates the initiation of transcription of the plc gene, possibly through direct contact with RNA polymerase.

Keywords: curved DNA, gene expression, Clostridium perfringens, phospholipase C




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