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Journal of Bacteriology, March 2000, p. 1313-1320, Vol. 182, No. 5
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Partitioning of the Linear Chromosome during Sporulation of Streptomyces coelicolor A3(2) Involves an oriC-Linked parAB Locus

Hyun-Jin Kim,1,dagger Michael J. Calcutt,2 Francis J. Schmidt,3 and Keith F. Chater1,*

John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, United Kingdom,1 and Department of Molecular Microbiology and Immunology, University of Missouri---Columbia and the Cancer Research Center,2 and Department of Biochemistry, University of Missouri---Columbia,3 Columbia, Missouri

Received 23 September 1999/Accepted 13 December 1999

Candidate partitioning genes (parA and parB) for the linear chromosome of Streptomyces coelicolor were identified by DNA sequencing in a series of seven genes located between rnpA and trxA near the chromosomal replication origin. The most likely translation start point of parB overlapped the parA stop codon, suggestive of coregulation, and transcription analysis suggested that the two genes formed an operon. Deletion of part of parB had no effect on the growth or appearance of colonies but caused a deficiency in DNA partitioning during the multiple septation events involved in converting aerial hyphae into long chains of spores. At least 13% of spore compartments failed to inherit the normal DNA allocation. The same phenotype was obtained with a deletion removing a segment of DNA from both parA and parB. Reinforcing the idea of a special role for the par locus during sporulation, the stronger of two parAB promoters was greatly upregulated at about the time when sporulation septation was maximal in colonies. Three copies of a 14-bp inverted repeat (GTTTCACGTGAAAC) were found in or near the parAB genes, and at least 12 more identical copies were identified within 100 kb of oriC from the growing genome sequence database. Only one perfect copy of the 14-bp sequence was present in approximately 5 Mb of sequence available from the rest of the genome. The 14-bp sequence was similar to sequences identified as binding sites for Spo0J, a ParB homologue from Bacillus subtilis believed to be important for DNA partitioning (D. C.-H. Lin and A. D. Grossman, Cell 92:675-685, 1998). One of these sites encompassed the transcription start point of the stronger parA promoter.

* Corresponding author. Mailing address: Department of Genetics, John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, United Kingdom. Phone: 44 (0)1603 452571. Fax: 44 (0)1603 456844. E-mail: chater{at}bbsrc.ac.uk.

dagger Present address: Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA 02111-1800.

Journal of Bacteriology, March 2000, p. 1313-1320, Vol. 182, No. 5
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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