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06 16, 2020;10 (1): 9694.

A role of the Nse4 kleisin and Nse1/Nse3 KITE subunits in the ATPase cycle of SMC5/6.

Lucie Vondrova, Peter Kolesar, Marek Adamus, Matej Nociar, Antony W Oliver, Jan J Palecek
Author Information
  1. Lucie Vondrova: National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kamenice 5, 62500, Brno, Czech Republic.
  2. Peter Kolesar: National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kamenice 5, 62500, Brno, Czech Republic.
  3. Marek Adamus: Mendel Centre for Plant Genomics and Proteomics, Central European Institute of Technology, Masaryk University, Kamenice 5, 62500, Brno, Czech Republic.
  4. Matej Nociar: National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kamenice 5, 62500, Brno, Czech Republic.
  5. Antony W Oliver: Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Falmer, Brighton, BN1 9RQ, United Kingdom.
  6. Jan J Palecek: National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kamenice 5, 62500, Brno, Czech Republic. jpalecek@sci.muni.cz.
PMID: 32546830 DOI: 10.1038/s41598-020-66647-w

Abstract

The SMC (Structural Maintenance of Chromosomes) complexes are composed of SMC dimers, kleisin and kleisin-interacting (HAWK or KITE) subunits. Mutual interactions of these subunits constitute the basal architecture of the SMC complexes. In addition, binding of ATP molecules to the SMC subunits and their hydrolysis drive dynamics of these complexes. Here, we developed new systems to follow the interactions between SMC5/6 subunits and the relative stability of the complex. First, we show that the N-terminal domain of the Nse4 kleisin molecule binds to the SMC6 neck and bridges it to the SMC5 head. Second, binding of the Nse1 and Nse3 KITE proteins to the Nse4 linker increased stability of the ATP-free SMC5/6 complex. In contrast, binding of ATP to SMC5/6 containing KITE subunits significantly decreased its stability. Elongation of the Nse4 linker partially suppressed instability of the ATP-bound complex, suggesting that the binding of the KITE proteins to the Nse4 linker constrains its limited size. Our data suggest that the KITE proteins may shape the Nse4 linker to fit the ATP-free complex optimally and to facilitate opening of the complex upon ATP binding. This mechanism suggests an important role of the KITE subunits in the dynamics of the SMC5/6 complexes.

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Grants

  1. MR/P018955/1/Medical Research Council

MeSH Term

Adenosine Triphosphatases
Carrier Proteins
Cell Cycle Proteins
Macromolecular Substances
Mutagenesis, Site-Directed
Nuclear Proteins
Protein Binding
Schizosaccharomyces
Schizosaccharomyces pombe Proteins
Sequence Alignment
Two-Hybrid System Techniques

Chemicals

Carrier Proteins
Cell Cycle Proteins
Macromolecular Substances
Nse1 protein, S pombe
Nse3 protein, S pombe
Nse4 protein, S pombe
Nuclear Proteins
Schizosaccharomyces pombe Proteins
Smc5 protein, S pombe
smc6 protein, S pombe
Adenosine Triphosphatases
Journal Article Research Support, Non-U.S. Gov't

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