Cyclins B1, T1, and H differ in their molecular mode of interaction with cytomegalovirus protein kinase pUL97

Steingruber M, Keller L, Socher E, Ferre S, Hesse AM, Coute Y, Hahn F, Buescher N, Plachter B, Sticht H, Marschall M (2019)

Publication Type: Journal article

Publication year: 2019

Journal

Book Volume: 294

Pages Range: 6188-6203

Journal Issue: 15

DOI: 10.1074/jbc.RA118.007049

Abstract

Human cytomegalovirus (HCMV) is a common -herpesvirus causing life-long latent infections. HCMV replication interferes with cell cycle regulation in host cells because the HCMV-encoded cyclin-dependent kinase (CDK) ortholog pUL97 extensively phosphorylates the checkpoint regulator retinoblastoma protein. pUL97 also interacts with cyclins B1, T1, and H, and recent findings have strongly suggested that these interactions influence pUL97 substrate recognition. Interestingly, here we detected profound mechanistic differences among these pUL97– cyclin interactions. Our study revealed the following. (i) pUL97 interacts with cyclins B1 and H in a manner dependent on pUL97 activity and HCMV-specific cyclin modulation, respectively. (ii) The phosphorylated state of both proteins is an important determinant of the pUL97–cyclin B1 interaction. (iii) Activated phospho-Thr-315 cyclin H is up-regulated during HCMV replication. (iv) Thr-315 phosphorylation is independent of intracellular pUL97 or CDK7 activity. (v) pUL97-mediated in vitro phosphorylation is detectable for cyclin B1 but not H. (vi) Mutual transphosphorylation between pUL97 and CDK7 is not detectable, and an MS-based phosphosite analysis indicated that pUL97 might unexpectedly not be phosphorylated in its T-loop. (vii) The binary complexes pUL97– cyclin H and CDK7– cyclin H as well as the ternary complex pUL97– cyclin-H–CDK7 are detectable in an assembly-based CoIP approach. (viii) pUL97 self-interaction can be bridged by the transcriptional cyclins T1 or H but not by the classical cell cycle–regulating B1 cyclin. Combined, our findings unravel a number of cyclin type–specific differences in pUL97 interactions and suggest a multifaceted regulatory impact of cyclins on HCMV replication.

Authors with CRIS profile

Involved external institutions

University of Grenoble Alpes (UGA) / Université de Grenoble France (FR) Johannes Gutenberg-Universität Mainz (JGU) Germany (DE)

How to cite

APA:

Steingruber, M., Keller, L., Socher, E., Ferre, S., Hesse, A.-M., Coute, Y.,... Marschall, M. (2019). Cyclins B1, T1, and H differ in their molecular mode of interaction with cytomegalovirus protein kinase pUL97. Journal of Biological Chemistry, 294(15), 6188-6203. https://dx.doi.org/10.1074/jbc.RA118.007049

MLA:

Steingruber, Mirjam, et al. "Cyclins B1, T1, and H differ in their molecular mode of interaction with cytomegalovirus protein kinase pUL97." Journal of Biological Chemistry 294.15 (2019): 6188-6203.

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