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Identification of Yin-Yang regulators and a phosphorylation consensus for male germ cell-associated kinase (MAK)-related kinase

Fu, Zheng ; Larson, Katherine A. ; Chitta, Raghu K. ; Parker, Sirlester A. ; Turk, Benjamin E. ; Lawrence, Matthew W. ; Kaldis, Philipp LU orcid ; Galaktionov, Konstantin ; Cohn, Steven M. and Shabanowitz, Jeffrey , et al. (2006) In Molecular and Cellular Biology 26(22). p.8639-8654
Abstract

MAK (male germ cell-associated protein kinase) and MRK/ICK (MAK-related kinase/intestinal cell kinase) are human homologs of Ime2p in Saccharomyces cerevisiae and of Mde3 and Pit1 in Schizosaccharomyces pombe and are similar to human cyclin-dependent kinase 2 (CDK2) and extracellular signal-regulated kinase 2 (ERK2). MAK and MRK require dual phosphorylation in a TDY motif catalyzed by an unidentified human threonine kinase and tyrosine autophosphorylation. Herein, we establish that human CDK-related kinase CCRK (cell cycle-related kinase) is an activating T157 kinase for MRK, whereas active CDIC7/cyclin H/MAT1 complexes phosphorylate CDK2 but not MRK. Protein phosphatase 5 (PP5) interacts with MRK in a complex and dephosphorylates MRK... (More)

MAK (male germ cell-associated protein kinase) and MRK/ICK (MAK-related kinase/intestinal cell kinase) are human homologs of Ime2p in Saccharomyces cerevisiae and of Mde3 and Pit1 in Schizosaccharomyces pombe and are similar to human cyclin-dependent kinase 2 (CDK2) and extracellular signal-regulated kinase 2 (ERK2). MAK and MRK require dual phosphorylation in a TDY motif catalyzed by an unidentified human threonine kinase and tyrosine autophosphorylation. Herein, we establish that human CDK-related kinase CCRK (cell cycle-related kinase) is an activating T157 kinase for MRK, whereas active CDIC7/cyclin H/MAT1 complexes phosphorylate CDK2 but not MRK. Protein phosphatase 5 (PP5) interacts with MRK in a complex and dephosphorylates MRK at T157 in vitro and in situ. Thus, CCRK and PP5 are yin-yang regulators of T157 phosphorylation. To determine a substrate consensus, we screened a combinatorial peptide library with active MRK. MRK preferentially phosphorylates R-P-X-S/T-P sites, with the preference for arginine at position -3 (P-3) being more stringent than for prolines at P-2 and P+1. Using the consensus, we identified a putative phosphorylation site (RPLT
1080S) for MRK in human Scythe, an antiapoptotic protein that interacts with MRK. MRK phosphorylates Scythe at T1080 in vitro as determined by site-directed mutagenesis and mass spectrometry, supporting the consensus and suggesting Scythe as a physiological substrate for MRK.

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publishing date
type
Contribution to journal
publication status
published
in
Molecular and Cellular Biology
volume
26
issue
22
pages
8639 - 8654
publisher
American Society for Microbiology
external identifiers
  • pmid:16954377
  • scopus:33751017086
ISSN
0270-7306
DOI
10.1128/MCB.00816-06
language
English
LU publication?
no
id
9ab3f070-39f7-4121-a99d-206ef8190db8
date added to LUP
2019-09-18 14:21:37
date last changed
2024-09-05 08:58:31
@article{9ab3f070-39f7-4121-a99d-206ef8190db8,
  abstract     = {{<p>MAK (male germ cell-associated protein kinase) and MRK/ICK (MAK-related kinase/intestinal cell kinase) are human homologs of Ime2p in Saccharomyces cerevisiae and of Mde3 and Pit1 in Schizosaccharomyces pombe and are similar to human cyclin-dependent kinase 2 (CDK2) and extracellular signal-regulated kinase 2 (ERK2). MAK and MRK require dual phosphorylation in a TDY motif catalyzed by an unidentified human threonine kinase and tyrosine autophosphorylation. Herein, we establish that human CDK-related kinase CCRK (cell cycle-related kinase) is an activating T157 kinase for MRK, whereas active CDIC7/cyclin H/MAT1 complexes phosphorylate CDK2 but not MRK. Protein phosphatase 5 (PP5) interacts with MRK in a complex and dephosphorylates MRK at T157 in vitro and in situ. Thus, CCRK and PP5 are yin-yang regulators of T157 phosphorylation. To determine a substrate consensus, we screened a combinatorial peptide library with active MRK. MRK preferentially phosphorylates R-P-X-S/T-P sites, with the preference for arginine at position -3 (P-3) being more stringent than for prolines at P-2 and P+1. Using the consensus, we identified a putative phosphorylation site (RPLT<br>
                        <sup>1080</sup>S) for MRK in human Scythe, an antiapoptotic protein that interacts with MRK. MRK phosphorylates Scythe at T1080 in vitro as determined by site-directed mutagenesis and mass spectrometry, supporting the consensus and suggesting Scythe as a physiological substrate for MRK.</p>}},
  author       = {{Fu, Zheng and Larson, Katherine A. and Chitta, Raghu K. and Parker, Sirlester A. and Turk, Benjamin E. and Lawrence, Matthew W. and Kaldis, Philipp and Galaktionov, Konstantin and Cohn, Steven M. and Shabanowitz, Jeffrey and Hunt, Donald F. and Sturgill, Thomas W.}},
  issn         = {{0270-7306}},
  language     = {{eng}},
  month        = {{11}},
  number       = {{22}},
  pages        = {{8639--8654}},
  publisher    = {{American Society for Microbiology}},
  series       = {{Molecular and Cellular Biology}},
  title        = {{Identification of Yin-Yang regulators and a phosphorylation consensus for male germ cell-associated kinase (MAK)-related kinase}},
  url          = {{http://dx.doi.org/10.1128/MCB.00816-06}},
  doi          = {{10.1128/MCB.00816-06}},
  volume       = {{26}},
  year         = {{2006}},
}