A novel type of DNA curvature present in a Clostridium perfringens ferredoxin gene: Characterization and role in gene expression

Masato Kaji, Osamu Matsushita, Eiji Tamai, Shigeru Miyata, Yuki Taniguchi, Seiko Shimamoto, Seiichi Katayama, Shushi Morita, Akinobu Okabe

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Abstract

This study has revealed that a Clostridium perfringens ferredoxin gene (per-fdx) possesses a novel type of DNA curvature, which is formed by five phased A-tracts extending from upstream to downstream of the -35 region. The three A-tracts upstream of the promoter and the two within the promoter are located at the positions corresponding to A-tracts present in a C. perfringens phospholipase C gene (plc) and a Clostridium pasteurianum ferredoxin gene (pas-fdx), respectively. DNA fragments of the per-fdx, pas-fdx and plc genes (nucleotide positions -69 to + 1 relative to the transcription initiation site) were fused to a chloramphenicol acetyltransferase reporter gene on a plasmid, pPSV, and their in vivo promoter activities were examined by assaying the chloramphenicol acetyltransferase activity of each C. perfringens transformant. Comparison of the three constructs showed that the order of promoter activity is, in descending order, per-fdx, pas-fdx and plc. Deletion of the three upstream A-tracts of the per-fdx gene drastically decreased the promoter activity, as demonstrated previously for the plc promoter. Substitution of the most downstream A-tract decreased the promoter activities of the per-fdx and pas-fdx genes. These results indicate that not only the phased A-tracts upstream of the promoter but also those within the promoter stimulate the promoter activity, and suggest that the high activity of the per-fdx promoter is due to the combined effects of these two types of A-tracts.

Original languageEnglish
Pages (from-to)3083-3091
Number of pages9
JournalMicrobiology
Volume149
Issue number11
DOIs
Publication statusPublished - Nov 2003

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ASJC Scopus subject areas

  • Microbiology

Cite this

Kaji, M., Matsushita, O., Tamai, E., Miyata, S., Taniguchi, Y., Shimamoto, S., Katayama, S., Morita, S., & Okabe, A. (2003). A novel type of DNA curvature present in a Clostridium perfringens ferredoxin gene: Characterization and role in gene expression. Microbiology, 149(11), 3083-3091. https://doi.org/10.1099/mic.0.26503-0