SMC Condensation Centers in Bacillus subtilis Are Dynamic Structures

Author:

Kleine Borgmann Luise A. K.1,Hummel Hanna1,Ulbrich Maximilian H.23,Graumann Peter L.134

Affiliation:

1. Microbiology, Faculty of Biology, University of Freiburg, Freiburg, Germany

2. Faculty of Medicine, Institute of Physiology II, Albert Ludwigs Universität Freiburg, Freiburg, Germany

3. BIOSS Centre for Biological Signalling Studies, Albert Ludwigs Universität Freiburg, Freiburg, Germany

4. Synmikro, LOEWE-Zentrum für Synthetische Mikrobiologie, Marburg, Germany

Abstract

ABSTRACT SMC and MukB complexes consist of a central SMC dimer and two essential binding partners, ScpA and ScpB (MukE and MukF), and are crucial for correct chromosome compaction and segregation. The complexes form two bipolar assemblies on the chromosome, one in each cell half. Using fluorescence recovery after photobleaching (FRAP), we provide evidence that the SMC complex has high exchange rates. This depends to a considerable degree on de novo protein synthesis, revealing that the bacterial SMC complex has high on and off rates for binding to the chromosome. A mutation in SMC that affects ATPase activity and results in exaggerated DNA binding in vitro causes a strong segregation defect in vivo and affects the localization of the entire SMC complex, which localizes to many more sites in the cell than under normal conditions. These data indicate that ATP turnover is important for the function of Bacillus subtilis SMC. In contrast, the centromere protein Spo0J and DNA gyrase showed much less exchange between distinct binding sites on the chromosome than that seen with SMC. Binding of Spo0J to the origin regions was rather static and remained partially conserved until the next cell cycle. Our experiments reveal that the SMC complex has a high, condensin-like turnover rate and that an alteration of the ATPase cycle affects SMC function in vivo , while several nucleoid-associated proteins feature limited or slow exchange between different sites on the nucleoid, which may be the basis for epigenetic-like phenomena observed in bacteria.

Publisher

American Society for Microbiology

Subject

Molecular Biology,Microbiology

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