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J Mol Biol. 2019 May 17;431(11):2127-2142. doi: 10.1016/j.jmb.2019.04.005. Epub 2019 Apr 9.

Allosteric Regulation of Cyclin-B Binding by the Charge State of Catalytic Lysine in CDK1 Is Essential for Cell-Cycle Progression.

Author information

1
Department of Biological Sciences, Tata Institute of Fundamental Research (TIFR), Mumbai 400005, India.
2
Department of Chemical Sciences, Tata Institute of Fundamental Research (TIFR), Mumbai 400005, India.
3
Institut de Pharmacologie et de Biologie Structurale (IPBS), Toulouse 31400, France.
4
Laboratory of Cell Signalling, Centre for DNA Fingerprinting and Diagnostics (CDFD), Hyderabad 500039, India; Graduate Studies, Manipal Academy of Higher Education, Manipal 576104, India.
5
Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research (JNCASR), Bengaluru 560064, India.
6
Laboratory of Cell Signalling, Centre for DNA Fingerprinting and Diagnostics (CDFD), Hyderabad 500039, India.
7
Department of Chemical Sciences, Tata Institute of Fundamental Research (TIFR), Mumbai 400005, India. Electronic address: ravi.venkatramani@tifr.res.in.
8
Department of Biological Sciences, Tata Institute of Fundamental Research (TIFR), Mumbai 400005, India. Electronic address: ullas@tifr.res.in.

Abstract

Cyclin-dependent kinase 1 (CDK1) is essential for cell-cycle progression. While dependence of CDK activity on cyclin levels is well established, molecular mechanisms that regulate their binding are less understood. Here, we report for the first time that CDK1:cyclin-B binding is not default but rather determined by the evolutionarily conserved catalytic residue, lysine-33 in CDK1. We demonstrate that the charge state of this lysine allosterically remodels the CDK1:cyclin-B interface. Cell cycle-dependent acetylation of lysine-33 or its mutation to glutamine, which mimics acetylation, abrogates cyclin-B binding. Using biochemical approaches and atomistic molecular dynamics simulations, we have uncovered both short-range and long-range effects of perturbing the charged state of the catalytic lysine, which lead to inhibition of kinase activity. Specifically, although loss of the charge state of catalytic lysine did not impact ATP binding significantly, it altered its orientation in the active site. In addition, the catalytic lysine also acts as an intra-molecular electrostatic tether at the active site to orient structural elements interfacing with cyclin-B. Physiologically, opposing activities of SIRT1 and P300 regulate acetylation and thus control the charge state of lysine-33. Importantly, cells expressing acetylation mimic mutant of Cdc2/CDK1 in yeast are arrested in G2 and fail to divide, indicating the requirement of the deacetylated state of the catalytic lysine for cell division. Thus, by illustrating the molecular role of the catalytic lysine and cell cycle-dependent deacetylation as a determinant of CDK1:cyclin-B interaction, our results redefine the current model of CDK1 activation and cell-cycle progression.

KEYWORDS:

G2-M progression; PSTAIRE-helix; SIRT1; deacetylation; kinase activity

PMID:
30974121
DOI:
10.1016/j.jmb.2019.04.005

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