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Applied and Environmental Microbiology, June 2002, p. 2924-2933, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.2924-2933.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

oriC Region and Replication Termination Site, dif, of the Xanthomonas campestris pv. campestris 17 Chromosome

Ming-Ren Yen,¹ Nien-Tsung Lin,² Chih-Hsin Hung,¹ Ka-Tim Choy,¹ Shu-Fen Weng,¹ and Yi-Hsiung Tseng^1*

Institute of Molecular Biology, National Chung Hsing University, Taichung 402,¹ Department of Microbiology, Tzu-Chi University, Hualien 907, Taiwan, Republic of China²

Received 11 December 2001/ Accepted 2 April 2002

A 13-kb DNA fragment containing oriC and the flanking genes thdF, orf900, yidC, rnpA, rpmH, oriC, dnaA, dnaN, recF, and gyrB was cloned from the gram-negative plant pathogen Xanthomonas campestris pv. campestris 17. These genes are conserved in order with other eubacterial oriC genes and code for proteins that share high degrees of identity with their homologues, except for orf900, which has a homologue only in Xylella fastidiosa. The dnaA/dnaN intergenic region (273 bp) identified to be the minimal oriC region responsible for autonomous replication has 10 pure AT clusters of four to seven bases and only three consensus DnaA boxes. These findings are in disagreement with the notion that typical oriCs contain four or more DnaA boxes located upstream of the dnaA gene. The X. campestris pv. campestris 17 attB site required for site-specific integration of cloned fragments from filamentous phage Lf replicative form DNA was identified to be a dif site on the basis of similarities in nucleotide sequence and function with the Escherichia coli dif site required for chromosome dimer resolution and whose deletion causes filamentation of the cells. The oriC and dif sites were located at 12:00 and 6:00, respectively, on the circular X. campestris pv. campestris 17 chromosome map, similar to the locations found for E. coli sites. Computer searches revealed the presence of both the dif site and XerC/XerD recombinase homologues in 16 of the 42 fully sequenced eubacterial genomes, but eight of the dif sites are located far away from the 6:00 point instead of being placed opposite the cognate oriC. The differences in the relative position suggest that mechanisms different from that of E. coli may participate in the control of chromosome replication.

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* Corresponding author. Mailing address: Institute of Molecular Biology, National Chung Hsing University, Taichung 402, Taiwan, Republic of China. Phone: 886-4-2285-1885. Fax: 886-4-2287-4879. E-mail: yhtseng{at}dragon.nchu.edu.tw.

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Applied and Environmental Microbiology, June 2002, p. 2924-2933, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.2924-2933.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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