A novel regulatory motif at the hinge dimer interface of the MksB mediates dimerization and DNA binding activity

Pratibha, Kumari and Vinayak, Hegde and Anant, Bakshi and Suguna, M. and Dharma Prasad, M. and Gnanesh Kumar, B. S. and Dandamudi, Usharani and Kunal, Sharan and Ravi, Kumar (2025) A novel regulatory motif at the hinge dimer interface of the MksB mediates dimerization and DNA binding activity. Biochimie, 230. pp. 68-85.

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Abstract

The Structural Maintenance of Chromosome (SMC) protein is essential for various cellular processes, including chromosome organization, DNA repair, and genome stability. MksB, an alternative SMC protein present in prokaryotes, comprises a hinge dimerization domain and an ABC ATPase head domain linked by a coiled-coil arm. While hinge dimerization in bacterial and eukaryotic SMCs is attributed to conserved glycines, our study unveils the critical role of a novel KDDR motif located at a loop near the hinge dimer interface in Mycobacterium smegmatis MksB (MsMksB). We demonstrate the regulatory role of this motif in MsMksB dimerization and DNA binding activity. The K600D mutation in the KDDR motif induces MsMksB dimer-to-monomer conversion, highlighting the significance of this motif in MsMksB dimerization. Mass spectrometry-based mapping of the DNA binding site revealed the lysine's involvement in protein-DNA interaction. Monomers of the hinge domain lose DNA binding activity, and MsMksB single-arm mutants exhibit reduced DNA binding and ATPase activity, underscoring the importance of hinge-mediated dimerization in MsMksB function. Notably, the R603D mutant retains dimerization but shows compromised ATPase and DNA binding activities. Mutants with defective ATPase activity exhibit impaired DNA condensation in vivo. These findings provide novel regulatory insight into the mechanism of MksB dimerization and DNA binding, uncovering the fundamental processes of chromosome condensation and segregation.

Item Type:	Article
Uncontrolled Keywords:	MsMksB, Hinge, Dimerization, KDDR motif, DNA binding
Subjects:	500 Natural Sciences and Mathematics > 07 Life Sciences > 03 Biochemistry & Molecular Biology > 13 Molecular Biochemistry
Divisions:	Molecular Nutrition
Depositing User:	Somashekar K S
Date Deposited:	05 Mar 2025 10:32
Last Modified:	05 Mar 2025 10:32
URI:	http://ir.cftri.res.in/id/eprint/19181

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