Cell cycle-dependent phosphorylation of nucleophosmin and its potential regulation by peptidyl-prolyl cis/trans isomerase

Xuelian Zhao, Junfang Ji, Li-Rong Yu, Timothy Veenstra, Xin Wei Wang


Nucleophosmin (NPM) is a ubiquitously expressed phosphoprotein involved in many cellular processes. Phosphorylation is considered the major regulatory mechanism of the NPM protein, associated with diverse cellular events. In this study, we characterized the phosphorylation status of several physiological phosphorylation sites of NPM, especially the newly confirmed in vivo site threonine 95 (Thr95). NPM-Thr95 exhibits a transient and cell cycle-dependent phosphorylation state compared to several other in vivo phosphorylation sites examined, including Ser4, Thr199 and Thr234/Thr237. In addition, we characterized a functional interaction between NPM and the peptidyl-prolyl isomerase Pin1, which specifically bind to each other during mitosis. The demonstration of this binding represents a novel post-phosphorylation regulatory mechanism for NPM that has not been investigated before. Mutated Pin1 putative binding sites result in defected cell division and reduced number of mitotic cells, suggesting that post-phosphorylation is important for NPM in regulating cell cycle progression.


nucleophosmin; Pin1; cell cycle; phosphorylation


Budhu AS & Wang XW 2005 Loading and unloading: orchestrating centrosome duplication and spindle assembly by Ran/Crm1. Cell Cycle 4 1510-1514

Cha H, Hancock C, Dangi S, Maiguel D, Carrier F & Shapiro P 2004 Phosphorylation regulates nucleophosmin targeting to the centrosome during mitosis as detected by cross-reactive phosphorylation-specific MKK1/MKK2 antibodies. Biochem J 378 857-865

Chan N & Meng Lim T 2015 Cytoplasmic nucleophosmin has elevated T199 phosphorylation upon which G2/M phase progression is dependent. Sci Rep 5 11777

Davis FM, Tsao TY, Fowler SK & Rao PN 1983 Monoclonal antibodies to mitotic cells. Proc Natl Acad Sci U S A 80 2926-2930

Ganem NJ, Godinho SA & Pellman D 2009 A mechanism linking extra centrosomes to chromosomal instability. Nature 460 278-282

Hernandez-Verdun D 2011 Assembly and disassembly of the nucleolus during the cell cycle. Nucleus 2 189-194

Herrera JE, Savkur R & Olson MO 1995 The ribonuclease activity of nucleolar protein B23. Nucleic Acids Res 23 3974-3979

Hinchcliffe EH & Sluder G 2001 “It takes two to tango": understanding how centrosome duplication is regulated throughout the cell cycle. Genes Dev 15 1167-1181

Innes BT, Bailey ML, Brandl CJ, Shilton BH & Litchfield DW 2013 Non-catalytic participation of the Pin1 peptidyl-prolyl isomerase domain in target binding. Front Physiol 4 18

Lindstrom MS 2011 NPM1/B23: A Multifunctional Chaperone in Ribosome Biogenesis and Chromatin Remodeling. Biochem Res Int 2011 195209

Liou YC, Ryo A, Huang HK, Lu PJ, Bronson R, Fujimori F, Uchida T, Hunter T & Lu KP 2002 Loss of Pin1 function in the mouse causes phenotypes resembling cyclin D1-null phenotypes. Proc Natl Acad Sci U S A 99 1335-1340

Lu KP, Hanes SD & Hunter T 1996 A human peptidyl-prolyl isomerase essential for regulation of mitosis. Nature 380 544-547

Lu PJ, Zhou XZ, Shen M & Lu KP 1999 Function of WW domains as phosphoserine- or phosphothreonine-binding modules. Science 283 1325-1328

Maher AI, el-Kashlan HK, Soliman Y & Galal R 1990 Rhinoscleroma: management by carbon dioxide surgical laser. Laryngoscope 100 783-788

Nigg EA 2002 Centrosome aberrations: cause or consequence of cancer progression? Nat Rev Cancer 2 815-825

Nigg EA, Cajanek L & Arquint C 2014 The centrosome duplication cycle in health and disease. FEBS Lett 588 2366-2372

Nigg EA & Stearns T 2011 The centrosome cycle: Centriole biogenesis, duplication and inherent asymmetries. Nat Cell Biol 13 1154-1160

Okuda M, Horn HF, Tarapore P, Tokuyama Y, Smulian AG, Chan PK, Knudsen ES, Hofmann IA, Snyder JD, Bove KE & Fukasawa K 2000 Nucleophosmin/B23 is a target of CDK2/cyclin E in centrosome duplication. Cell 103 127-140

Okuwaki M 2008 The structure and functions of NPM1/Nucleophsmin/B23, a multifunctional nucleolar acidic protein. J Biochem 143 441-448

Ranganathan R, Lu KP, Hunter T & Noel JP 1997 Structural and functional analysis of the mitotic rotamase Pin1 suggests substrate recognition is phosphorylation dependent. Cell 89 875-886

Rizzolio F, Lucchetti C, Caligiuri I, Marchesi I, Caputo M, Klein-Szanto AJ, Bagella L, Castronovo M & Giordano A 2012 Retinoblastoma tumor-suppressor protein phosphorylation and inactivation depend on direct interaction with Pin1. Cell Death Differ 19 1152-1161

Silkworth WT, Nardi IK, Scholl LM & Cimini D 2009 Multipolar spindle pole coalescence is a major source of kinetochore mis-attachment and chromosome mis-segregation in cancer cells. PLoS One 4 e6564

Suizu F, Ryo A, Wulf G, Lim J & Lu KP 2006 Pin1 regulates centrosome duplication, and its overexpression induces centrosome amplification, chromosome instability, and oncogenesis. Mol Cell Biol 26 1463-1479

Ubersax JA & Ferrell Jr JE 2007 Mechanisms of specificity in protein phosphorylation. Nat Rev Mol Cell Biol 8 530-541

Wang W, Budhu A, Forgues M & Wang XW 2005 Temporal and spatial control of nucleophosmin by the Ran-Crm1 complex in centrosome duplication. Nat Cell Biol 7 823-830

Wei S, Kozono S, Kats L, Nechama M, Li W, Guarnerio J, Luo M, You MH, Yao Y, Kondo A, Hu H, Bozkurt G, Moerke NJ, Cao S, Reschke M, Chen CH, Rego EM, Lo-Coco F, Cantley LC, Lee TH, Wu H, Zhang Y, Pandolfi PP, Zhou XZ & Lu KP 2015 Active Pin1 is a key target of all-trans retinoic acid in acute promyelocytic leukemia and breast cancer. Nat Med 21 457-466

Yeh ES, Lew BO & Means AR 2006 The loss of PIN1 deregulates cyclin E and sensitizes mouse embryo fibroblasts to genomic instability. J Biol Chem 281 241-251

Yu LR, Zhu Z, Chan KC, Issaq HJ, Dimitrov DS & Veenstra TD 2007 Improved titanium dioxide enrichment of phosphopeptides from HeLa cells and high confident phosphopeptide identification by cross-validation of MS/MS and MS/MS/MS spectra. J Proteome Res 6 4150-4162

Zheng H, You H, Zhou XZ, Murray SA, Uchida T, Wulf G, Gu L, Tang X, Lu KP & Xiao ZX 2002 The prolyl isomerase Pin1 is a regulator of p53 in genotoxic response. Nature 419 849-853

Zhou W, Yang Q, Low CB, Karthik BC, Wang Y, Ryo A, Yao SQ, Yang D & Liou YC 2009 Pin1 catalyzes conformational changes of Thr-187 in p27Kip1 and mediates its stability through a polyubiquitination process. J Biol Chem 284 23980-23988

Zyss D & Gergely F 2009 Centrosome function in cancer: guilty or innocent? Trends Cell Biol 19 334-346

Full Text: PDF


  • There are currently no refbacks.

Copyright © 2017 Lorem Ipsum Press