New Cdc2 Tyr 4 Phosphorylation by DsRNA-activated Protein Kinase Triggers Cdc2 Polyubiquitination and G2 Arrest Under Genotoxic Stresses

Overview

Journal EMBO Rep

Specialty Molecular Biology

Date 2010 Apr 17

PMID 20395957

Citations 16

Authors

Cheol-Hee Yoon

Mohammad Alam Miah

Kwang Pyo Kim

Yong-Soo Bae

Affiliations

Soon will be listed here.

Abstract

Cell division cycle 2 (Cdc2) protein is an essential subunit of M-phase kinase (MPK), which has a key role in G2/M transition. Even though the control of MPK activity has been well established with regard to the phosphorylation of Cdc2 at Thr 14 and/or Tyr 15 and Thr 161, little is known about the proteolytic control of Cdc2. In this study, we observed that Cdc2 was downregulated under genotoxic stresses and that double-stranded RNA-activated protein kinase (PKR) was involved in the process. The PKR-mediated Tyr4 phosphorylation triggered Cdc2 ubiquitination. Phospho-mimic mutations at the Tyr 4 residue (Y4D or Y4E) caused significant ubiquitination of Cdc2 even in the absence of PKR. Our findings demonstrate that (i) PKR, Ser/Thr kinase, phosphorylates its new substrate Cdc2 at the Tyr 4 residue, (ii) PKR-mediated Tyr 4-phosphorylation facilitates Cdc2 ubiquitination and proteosomal degradation, (iii) unphosphorylated Tyr 4 prevents Cdc2 ubiquitination, and (iv) downstream from p53, PKR has a crucial role in G2 arrest and triggers Cdc2 downregulation under genotoxic conditions.

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References

Dagon Y, Dovrat S, Vilchik S, Hacohen D, Shlomo G, Sredni B . Double-stranded RNA-dependent protein kinase, PKR, down-regulates CDC2/cyclin B1 and induces apoptosis in non-transformed but not in v-mos transformed cells. Oncogene. 2002; 20(56):8045-56. DOI: 10.1038/sj.onc.1204945. View

Yoon C, Lee E, Lim D, Bae Y . PKR, a p53 target gene, plays a crucial role in the tumor-suppressor function of p53. Proc Natl Acad Sci U S A. 2009; 106(19):7852-7. PMC: 2683089. DOI: 10.1073/pnas.0812148106. View

Lee E, Lee M, Camus S, Ghim J, Yang M, Oh W . Differential regulation of p53 and p21 by MKRN1 E3 ligase controls cell cycle arrest and apoptosis. EMBO J. 2009; 28(14):2100-13. PMC: 2718286. DOI: 10.1038/emboj.2009.164. View

Baltzis D, Pluquet O, Papadakis A, Kazemi S, Qu L, Koromilas A . The eIF2alpha kinases PERK and PKR activate glycogen synthase kinase 3 to promote the proteasomal degradation of p53. J Biol Chem. 2007; 282(43):31675-87. DOI: 10.1074/jbc.M704491200. View

Raven J, Baltzis D, Wang S, Mounir Z, Papadakis A, Gao H . PKR and PKR-like endoplasmic reticulum kinase induce the proteasome-dependent degradation of cyclin D1 via a mechanism requiring eukaryotic initiation factor 2alpha phosphorylation. J Biol Chem. 2007; 283(6):3097-3108. DOI: 10.1074/jbc.M709677200. View

Su Q, Wang S, Baltzis D, Qu L, Wong A, Koromilas A . Tyrosine phosphorylation acts as a molecular switch to full-scale activation of the eIF2alpha RNA-dependent protein kinase. Proc Natl Acad Sci U S A. 2005; 103(1):63-8. PMC: 1324992. DOI: 10.1073/pnas.0508207103. View

Alisi A, Mele R, Spaziani A, Tavolaro S, Palescandolo E, Balsano C . Thr 446 phosphorylation of PKR by HCV core protein deregulates G2/M phase in HCC cells. J Cell Physiol. 2005; 205(1):25-31. DOI: 10.1002/jcp.20363. View

Taylor W, Stark G . Regulation of the G2/M transition by p53. Oncogene. 2001; 20(15):1803-15. DOI: 10.1038/sj.onc.1204252. View

Lee E, Yoon C, Kim Y, Bae Y . The double-strand RNA-dependent protein kinase PKR plays a significant role in a sustained ER stress-induced apoptosis. FEBS Lett. 2007; 581(22):4325-32. DOI: 10.1016/j.febslet.2007.08.001. View

10.

Gautier J, Solomon M, Booher R, Bazan J, Kirschner M . cdc25 is a specific tyrosine phosphatase that directly activates p34cdc2. Cell. 1991; 67(1):197-211. DOI: 10.1016/0092-8674(91)90583-k. View

11.

Kerr I, Brown R, Hovanessian A . Nature of inhibitor of cell-free protein synthesis formed in response to interferon and double-stranded RNA. Nature. 1977; 268(5620):540-2. DOI: 10.1038/268540a0. View

12.

Mueller P, Coleman T, Kumagai A, Dunphy W . Myt1: a membrane-associated inhibitory kinase that phosphorylates Cdc2 on both threonine-14 and tyrosine-15. Science. 1995; 270(5233):86-90. DOI: 10.1126/science.270.5233.86. View

13.

Wu R, Feng Q, Lonard D, OMalley B . SRC-3 coactivator functional lifetime is regulated by a phospho-dependent ubiquitin time clock. Cell. 2007; 129(6):1125-40. DOI: 10.1016/j.cell.2007.04.039. View

14.

Labbe J, Lee M, Nurse P, Picard A, Doree M . Activation at M-phase of a protein kinase encoded by a starfish homologue of the cell cycle control gene cdc2+. Nature. 1988; 335(6187):251-4. DOI: 10.1038/335251a0. View

15.

Watanabe N, Arai H, Nishihara Y, Taniguchi M, Watanabe N, Hunter T . M-phase kinases induce phospho-dependent ubiquitination of somatic Wee1 by SCFbeta-TrCP. Proc Natl Acad Sci U S A. 2004; 101(13):4419-24. PMC: 384762. DOI: 10.1073/pnas.0307700101. View

16.

Le Gac G, Esteve P, Ferec C, Pradhan S . DNA damage-induced down-regulation of human Cdc25C and Cdc2 is mediated by cooperation between p53 and maintenance DNA (cytosine-5) methyltransferase 1. J Biol Chem. 2006; 281(34):24161-70. DOI: 10.1074/jbc.M603724200. View