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Journal of Bacteriology, September 2006, p. 6184-6194, Vol. 188, No. 17
0021-9193/06/$08.00+0     doi:10.1128/JB.00528-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Cluster of DnaA Boxes Involved in Regulation of Streptomyces Chromosome Replication: from In Silico to In Vivo Studies

Aleksandra Smulczyk-Krawczyszyn,^1,2 Dagmara Jakimowicz,^1,2 Beata Ruban-Omiaowska,¹ Anna Zawilak-Pawlik,¹ Jerzy Majka,^1, Keith Chater,² and Jolanta Zakrzewska-Czerwiska^1*

Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, ul. Weigla 12, 53-114 Wrocaw, Poland,¹ John Innes Centre, Norwich Research Park, Colney, Norwich NR4 7UH, United Kingdom²

Received 13 April 2006/ Accepted 12 June 2006

In Streptomyces coelicolor, replication is initiated by the DnaA protein in the centrally located oriC region and proceeds bidirectionally until the replication forks reach the ends of the linear chromosome. We identified three clusters of DnaA boxes (H69, H24, and D78) which are in a relatively short segment of the chromosome centered on the oriC region. Of the clusters analyzed, D78 exhibited the highest affinity for the DnaA protein; the affinity of DnaA for the D78 cluster was about eightfold higher than the affinity for oriC. The high-affinity DnaA boxes appear to be involved in the control of chromosome replication. Deletion of D78 resulted in more frequent chromosome replication (an elevated ratio of origins to chromosome ends was observed) and activated aerial mycelium formation, leading to earlier colony maturation. In contrast, extra copies of D78 (delivered on a plasmid) caused slow colony growth, presumably because of a reduction in the frequency of initiation of chromosome replication. This suggests that the number of high-affinity DnaA boxes is relatively constant in hyphal compartments and that deletion of D78 therefore permits an increased copy number of either the chromosomal origin region or a plasmid harboring the D78 cluster. This system conceivably influences the timing of decisions to initiate aerial mycelial formation and sporulation.

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* Corresponding author. Mailing address: Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, ul. Weigla 12, 53-114 Wrocaw, Poland. Phone: 48 71 3709948. Fax: 48 71 3371382. E-mail: zakrzew{at}iitd.pan.wroc.pl.

Supplemental material for this article may be found at http://jb.asm.org/.

Present address: Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Mo.

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Journal of Bacteriology, September 2006, p. 6184-6194, Vol. 188, No. 17
0021-9193/06/$08.00+0     doi:10.1128/JB.00528-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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