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Molecular and Cellular Biology, August 2003, p. 5638-5650, Vol. 23, No. 16
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.16.5638-5650.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

A Prokaryotic Condensin/Cohesin-Like Complex Can Actively Compact Chromosomes from a Single Position on the Nucleoid and Binds to DNA as a Ring-Like Structure{dagger}

A. Volkov,1 J. Mascarenhas,1 C. Andrei-Selmer,1 H. D. Ulrich,2 and P. L. Graumann1*

Biochemie, Fachbereich Chemie, Philipps-Universität Marburg,1 Max-Planck Institut für Terrestrische Mikrobiologie, Marburg, Germany2

Received 18 October 2002/ Returned for modification 22 November 2002/ Accepted 16 May 2003

We show that Bacillus subtilis SMC (structural maintenance of chromosome protein) localizes to discrete foci in a cell cycle-dependent manner. Early in the cell cycle, SMC moves from the middle of the cell toward opposite cell poles in a rapid and dynamic manner and appears to interact with different regions on the chromosomes during the cell cycle. SMC colocalizes with its interacting partners, ScpA and ScpB, and the specific localization of SMC depends on both Scp proteins, showing that all three components of the SMC complex are required for proper localization. Cytological and biochemical experiments showed that dimeric ScpB stabilized the binding of ScpA to the SMC head domains. Purified SMC showed nonspecific binding to double-stranded DNA, independent of Scp proteins or ATP, and was retained on DNA after binding to closed DNA but not to linear DNA. The SMC head domains and hinge region did not show strong DNA binding activity, suggesting that the coiled-coil regions in SMC mediate an association with DNA and that SMC binds to DNA as a ring-like structure. The overproduction of SMC resulted in global chromosome compaction, while SMC was largely retained in bipolar foci, suggesting that the SMC complex forms condensation centers that actively affect global chromosome compaction from a defined position on the nucleoid.

* Corresponding author. Mailing address: Biochemie, Fachbereich Chemie, Philipps-Universität Marburg, Hans-Meerwein-Straße, Marburg, Germany. Phone: 49(0)64212825539. Fax: 49(0)64212822191. E-mail: graumann{at}chemie.uni-marburg.de.

{dagger} This report is dedicated to Richard Losick in celebration of his 60th birthday.

Molecular and Cellular Biology, August 2003, p. 5638-5650, Vol. 23, No. 16
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.16.5638-5650.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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