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Journal of Bacteriology, February 1999, p. 923-933, Vol. 181, No. 3
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Gene Duplication and Multiplicity of Collagenases in Clostridium histolyticum

Osamu Matsushita,1 Chang-Min Jung,1 Seiichi Katayama,1 Junzaburo Minami,1 Yukie Takahashi,2 and Akinobu Okabe1,*

Department of Microbiology, Faculty of Medicine, Kagawa Medical University, Kagawa 761-0793,1 and Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, Fukuyama, Hiroshima 729-0292,2 Japan

Received 30 June 1998/Accepted 16 November 1998

Clostridium histolyticum collagenase contains a number of different active components. Previously we have shown that colH encodes a 116-kDa collagenase (ColH) and a 98-kDa gelatinase. We purified a different 116-kDa collagenase (ColG) from the culture supernatant and sequenced its gene (colG). We also identified four other gelatinases (105, 82, 78, and 67 kDa) and determined their N-terminal amino acid sequences, all of which coincided with that of either ColG or ColH. Hybridization experiments showed that each gene is present in a single copy and each gene is transcribed into a single mRNA. These results suggest that all the gelatinases are produced from the respective full-length collagenase by the proteolytic removal of C-terminal fragments. The substrate specificities of the enzymes suggest that colG and colH encode class I and class II enzymes, respectively. Analysis of their DNA locations by pulsed-field gel electrophoresis and nucleotide sequencing of their surrounding regions revealed that the two genes are located in different sites on the chromosome. C. histolyticum colG is more similar to C. perfringens colA than to colH in terms of domain structure. Both colG and colA have a homologous gene, mscL, at their 3' ends. These results suggest that gene duplication and segment duplication have occurred in an ancestor cell common to C. histolyticum and C. perfringens and that further divergence of the parent gene produced colG and colA.


* Corresponding author. Mailing address: Department of Microbiology, Faculty of Medicine, Kagawa Medical University, 1750-1 Ikenobe, Miki-cho, Kagawa 761-0793, Japan. Phone: 81 (87) 891-2129. Fax: 81 (87) 898-7109. E-mail: microbio{at}kms.ac.jp.


Journal of Bacteriology, February 1999, p. 923-933, Vol. 181, No. 3
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



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