Nuclear HuR Accumulation Through Phosphorylation by Cdk1

Overview

Journal Genes Dev

Specialty Molecular Biology

Date 2008 Jul 3

PMID 18593881

Citations 127

Authors

Hyeon Ho Kim

Kotb Abdelmohsen

Ashish Lal

Rudolf Pullmann Jr

Xiaoling Yang

Stefanie Galban

Subramanya Srikantan

Jennifer L Martindale

Justin Blethrow

Kevan M Shokat

Myriam Gorospe

Affiliations

Soon will be listed here.

Abstract

A predominantly nuclear RNA-binding protein, HuR translocates to the cytoplasm in response to stress and proliferative signals, where it stabilizes or modulates the translation of target mRNAs. Here, we present evidence that HuR phosphorylation at S202 by the G2-phase kinase Cdk1 influences its subcellular distribution. HuR was specifically phosphorylated in synchronous G2-phase cultures; its cytoplasmic levels increased by Cdk1-inhibitory interventions and declined in response to Cdk1-activating interventions. In keeping with the prominently cytoplasmic location of the nonphosphorylatable point mutant HuR(S202A), phospho-HuR(S202) was shown to be predominantly nuclear using a novel anti-phospho-HuR(S202) antibody. The enhanced cytoplasmic presence of unphosphorylated HuR was linked to its decreased association with 14-3-3 and to its heightened binding to target mRNAs. Our findings suggest that Cdk1 phosphorylates HuR during G2, thereby helping to retain it in the nucleus in association with 14-3-3 and hindering its post-transcriptional function and anti-apoptotic influence.

Citing Articles

Structural, biological, and biomedical implications of mRNA interactions with the master regulator HuR.

Clark M, Farinha A, Morrison A, Lisi G NAR Mol Med. 2025; 2(1):ugaf002.

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PMID: 38809616 PMC: 11377808. DOI: 10.1681/ASN.0000000000000382.

pADP-ribosylation regulates the cytoplasmic localization, cleavage, and pro-apoptotic function of HuR.

Ashour K, Sali S, Aldoukhi A, Hall D, Mubaid S, Busque S Life Sci Alliance. 2024; 7(6).

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References

Zhang W, Wagner B, Ehrenman K, Schaefer A, DeMaria C, Crater D . Purification, characterization, and cDNA cloning of an AU-rich element RNA-binding protein, AUF1. Mol Cell Biol. 1993; 13(12):7652-65. PMC: 364837. DOI: 10.1128/mcb.13.12.7652-7665.1993. View

Yu C, Rahmani M, Dai Y, Conrad D, Krystal G, Dent P . The lethal effects of pharmacological cyclin-dependent kinase inhibitors in human leukemia cells proceed through a phosphatidylinositol 3-kinase/Akt-dependent process. Cancer Res. 2003; 63(8):1822-33. View

Moore M . From birth to death: the complex lives of eukaryotic mRNAs. Science. 2005; 309(5740):1514-8. DOI: 10.1126/science.1111443. View

Chen C, Gherzi R, Ong S, Chan E, Raijmakers R, Pruijn G . AU binding proteins recruit the exosome to degrade ARE-containing mRNAs. Cell. 2001; 107(4):451-64. DOI: 10.1016/s0092-8674(01)00578-5. View

Gallouzi I, Steitz J . Delineation of mRNA export pathways by the use of cell-permeable peptides. Science. 2001; 294(5548):1895-901. DOI: 10.1126/science.1064693. View

Poon R, Jiang W, Toyoshima H, Hunter T . Cyclin-dependent kinases are inactivated by a combination of p21 and Thr-14/Tyr-15 phosphorylation after UV-induced DNA damage. J Biol Chem. 1996; 271(22):13283-91. DOI: 10.1074/jbc.271.22.13283. View

Blethrow J, Glavy J, Morgan D, Shokat K . Covalent capture of kinase-specific phosphopeptides reveals Cdk1-cyclin B substrates. Proc Natl Acad Sci U S A. 2008; 105(5):1442-7. PMC: 2234163. DOI: 10.1073/pnas.0708966105. View

Levy N, Chung S, Furneaux H, Levy A . Hypoxic stabilization of vascular endothelial growth factor mRNA by the RNA-binding protein HuR. J Biol Chem. 1998; 273(11):6417-23. DOI: 10.1074/jbc.273.11.6417. View

Johnson B, Stehn J, Yaffe M, Blackwell T . Cytoplasmic localization of tristetraprolin involves 14-3-3-dependent and -independent mechanisms. J Biol Chem. 2002; 277(20):18029-36. DOI: 10.1074/jbc.M110465200. View

10.

Keene J . Why is Hu where? Shuttling of early-response-gene messenger RNA subsets. Proc Natl Acad Sci U S A. 1999; 96(1):5-7. PMC: 33538. DOI: 10.1073/pnas.96.1.5. View

11.

Tran H, Brunet A, Grenier J, Datta S, Fornace Jr A, DiStefano P . DNA repair pathway stimulated by the forkhead transcription factor FOXO3a through the Gadd45 protein. Science. 2002; 296(5567):530-4. DOI: 10.1126/science.1068712. View

12.

Lal A, Abdelmohsen K, Pullmann R, Kawai T, Galban S, Yang X . Posttranscriptional derepression of GADD45alpha by genotoxic stress. Mol Cell. 2006; 22(1):117-28. DOI: 10.1016/j.molcel.2006.03.016. View

13.

Nabors L, Gillespie G, Harkins L, King P . HuR, a RNA stability factor, is expressed in malignant brain tumors and binds to adenine- and uridine-rich elements within the 3' untranslated regions of cytokine and angiogenic factor mRNAs. Cancer Res. 2001; 61(5):2154-61. View

14.

Lal A, Mazan-Mamczarz K, Kawai T, Yang X, Martindale J, Gorospe M . Concurrent versus individual binding of HuR and AUF1 to common labile target mRNAs. EMBO J. 2004; 23(15):3092-102. PMC: 514922. DOI: 10.1038/sj.emboj.7600305. View

15.

Gherzi R, Trabucchi M, Ponassi M, Ruggiero T, Corte G, Moroni C . The RNA-binding protein KSRP promotes decay of beta-catenin mRNA and is inactivated by PI3K-AKT signaling. PLoS Biol. 2006; 5(1):e5. PMC: 1702562. DOI: 10.1371/journal.pbio.0050005. View

16.

Wang W, Yang X, Kawai T, Lopez de Silanes I, Mazan-Mamczarz K, Chen P . AMP-activated protein kinase-regulated phosphorylation and acetylation of importin alpha1: involvement in the nuclear import of RNA-binding protein HuR. J Biol Chem. 2004; 279(46):48376-88. DOI: 10.1074/jbc.M409014200. View

17.

Mitchell P, Tollervey D . mRNA stability in eukaryotes. Curr Opin Genet Dev. 2000; 10(2):193-8. DOI: 10.1016/s0959-437x(00)00063-0. View

18.

Yaffe M . How do 14-3-3 proteins work?-- Gatekeeper phosphorylation and the molecular anvil hypothesis. FEBS Lett. 2002; 513(1):53-7. DOI: 10.1016/s0014-5793(01)03288-4. View

19.

van der Giessen K, Di-Marco S, Clair E, Gallouzi I . RNAi-mediated HuR depletion leads to the inhibition of muscle cell differentiation. J Biol Chem. 2003; 278(47):47119-28. DOI: 10.1074/jbc.M308889200. View

20.

Antic D, Lu N, Keene J . ELAV tumor antigen, Hel-N1, increases translation of neurofilament M mRNA and induces formation of neurites in human teratocarcinoma cells. Genes Dev. 1999; 13(4):449-61. PMC: 316478. DOI: 10.1101/gad.13.4.449. View