Abstract
Ret finger protein-like 1 (RFPL1) is a primate-specific target gene of Pax6, a key transcription factor for pancreas, eye and neocortex development. However, its cellular activity remains elusive. In this article, we report that Pax6-elicited expression of the human (h)RFPL1 gene in HeLa cells can be enhanced by in vivo p53 binding to its promoter and therefore investigated the hypothesis that hRFPL1 regulates cell-cycle progression. Upon expression in these cells, hRFPL1 decreased cell number through a kinase-dependent mechanism as PKC activates and Cdc2 inhibits hRFPL1 activity. hRFPL1 antiproliferative activity led to an increased cell population in G2/M phase and specific cyclin B1 and Cdc2 downregulations, which were precluded by a proteasome inhibitor. Specifically, cytoplasm-localized hRFPL1 prevented cyclin B1 and Cdc2 accumulation during interphase. Consequently, cells showed a delayed entry into mitosis and cell-cycle lengthening resulting from a threefold increase in G2 phase duration. Given previous reports that RFPL1 is expressed during cell differentiation, its impact on cell-cycle lengthening therefore provides novel insights into primate-specific development.
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Abbreviations
- cPFTα:
-
cyclic-Pifithrin-α
- RDM:
-
RFPL-defining motif
- RFPL:
-
Ret finger protein-like
- RSH:
-
RFPL1,2,3-specifying helix
- SNP:
-
single nucleotide polymorphism
References
Englund C, Fink A, Lau C, Pham D, Daza RA, Bulfone A et al. Pax6, Tbr2, and Tbr1 are expressed sequentially by radial glia, intermediate progenitor cells, and postmitotic neurons in developing neocortex. J Neurosci 2005; 25: 247–251.
Liu W, Lagutin OV, Mende M, Streit A, Oliver G . Six3 activation of Pax6 expression is essential for mammalian lens induction and specification. EMBO J 2006; 25: 5383–5395.
Mellitzer G, Bonné S, Luco RF, Van De Casteele M, Lenne-Samuel N, Collombat P et al. IA1 is NGN3-dependent and essential for differentiation of the endocrine pancreas. EMBO J 2006; 25: 1344–1352.
Bonnefont J, Nikolaev SI, Perrier AL, Guo S, Cartier L, Sorce S et al. Evolutionary forces shape the human RFPL1,2,3 genes toward a role in neocortex development. Am J Hum Genet 2008; 83: 208–218.
Rajkovic A, Lee JH, Yan C, Matzuk MM . The ret finger protein-like 4 gene, Rfpl4, encodes a putative E3 ubiquitin-protein ligase expressed in adult germ cells. Mech Dev 2002; 112: 173–177.
Suzumori N, Burns KH, Yan W, Matzuk MM . RFPL4 interacts with oocyte proteins of the ubiquitin–proteasome degradation pathway. Proc Natl Acad Sci USA 2003; 100: 550–555.
Ledan E, Polanski Z, Terret ME, Maro B . Meiotic maturation of the mouse oocyte requires an equilibrium between cyclin B synthesis and degradation. Dev Biol 2001; 232: 400–413.
Bassermann F, Peschel C, Duyster J . Mitotic entry: a matter of oscillating destruction. Cell Cycle 2005; 4: 1515–1517.
Cartier L, Laforge T, Feki A, Arnaudeau S, Dubois-Dauphin M, Krause KH . Pax6-induced alteration of cell fate: shape changes, expression of neuronal alpha tubulin, postmitotic phenotype, and cell migration. J Neurobiol 2006; 66: 421–436.
Komarov PG, Komarova EA, Kondratov RV, Christov-Tselkov K, Coon JS, Chernov MV et al. A chemical inhibitor of p53 that protects mice from the side effects of cancer therapy. Science 1999; 285: 1733–1737.
Wiznerowicz M, Trono D . Conditional suppression of cellular genes: lentivirus vector-mediated drug-inducible RNA interference. J Virol 2003; 77: 8957–8961.
Clement V, Dunand-Sauthier I, Wiznerowicz M, Clarkson SG . UV-induced apoptosis in XPG-deficient fibroblasts involves activation of CD95 and caspases but not p53. DNA Repair (Amst) 2007; 6: 602–614.
Sansom SN, Griffiths DS, Faedo A, Kleinjan DJ, Ruan Y, Smith J et al. The level of the transcription factor Pax6 is essential for controlling the balance between neural stem cell self-renewal and neurogenesis. PLoS Genet 2009; 5: e1000511.
Thomas M, Pim D, Banks L . The role of the E6–p53 interaction in the molecular pathogenesis of HPV. Oncogene 1999; 18: 7690–7700.
Lindqvist A, van Zon W, Karlsson Rosenthal C, Wolthuis RM . Cyclin B1-Cdk1 activation continues after centrosome separation to control mitotic progression. PLoS Biol 2007; 5: e123.
Solomon MJ, Glotzer M, Lee TH, Philippe M, Kirschner MW . Cyclin activation of p34cdc2. Cell 1990; 63: 1013–1024.
Yang J, Bardes ES, Moore JD, Brennan J, Powers MA, Kornbluth S . Control of cyclin B1 localization through regulated binding of the nuclear export factor CRM1. Genes Dev 1998; 12: 2131–2143.
Peters JM . The anaphase promoting complex/cyclosome: a machine designed to destroy. Nat Rev Mol Cell Biol 2006; 7: 644–656.
Hsieh YW, Yang XJ . Dynamic Pax6 expression during the neurogenic cell cycle influences proliferation and cell fate choices of retinal progenitors. Neural Dev 2009; 4: 32.
Heins N, Malatesta P, Cecconi F, Nakafuku M, Tucker KL, Hack MA et al. Glial cells generate neurons: the role of the transcription factor Pax6. Nat Neurosci 2002; 5: 308–315.
Suter DM, Tirefort D, Julien S, Krause KH . A Sox1 to Pax6 switch drives neuroectoderm to radial glia progression during differentiation of mouse embryonic stem cells. Stem Cells 2009; 27: 49–58.
Innocente SA, Abrahamson JL, Cogswell JP, Lee JM . p53 regulates a G2 checkpoint through cyclin B1. Proc Natl Acad Sci USA 1999; 96: 2147–2152.
Iwai H, Kim M, Yoshikawa Y, Ashida H, Ogawa M, Fujita Y et al. A bacterial effector targets Mad2L2, an APC inhibitor, to modulate host cell cycling. Cell 2007; 130: 611–623.
Yuan J, Krämer A, Matthess Y, Yan R, Spänkuch B, Gätje R et al. Stable gene silencing of cyclin B1 in tumor cells increases susceptibility to taxol and leads to growth arrest in vivo. Oncogene 2006; 25: 1753–1762.
Xie XH, An HJ, Kang S, Hong S, Choi YP, Kim YT et al. Loss of cyclin B1 followed by downregulation of cyclin A/Cdk2, apoptosis and antiproliferation in HeLa cell line. Int J Cancer 2005; 116: 520–525.
Ubersax JA, Woodbury EL, Quang PN, Paraz M, Blethrow JD, Shah K et al. Targets of the cyclin-dependent kinase Cdk1. Nature 2003; 425: 859–864.
Santamaria D, Barrière C, Cerqueira A, Hunt S, Tardy C, Newton K et al. Cdk1 is sufficient to drive the mammalian cell cycle. Nature 2007; 448: 811–815.
Listovsky T, Zor A, Laronne A, Brandeis M . Cdk1 is essential for mammalian cyclosome/APC regulation. Exp Cell Res 2000; 255: 184–191.
Menendez D, Krysiak O, Inga A, Krysiak B, Resnick MA, Schönfelder G . A SNP in the flt-1 promoter integrates the VEGF system into the p53 transcriptional network. Proc Natl Acad Sci USA 2006; 103: 1406–1411.
Menendez D, Inga A, Snipe J, Krysiak O, Schönfelder G, Resnick MA . A single-nucleotide polymorphism in a half-binding site creates p53 and estrogen receptor control of vascular endothelial growth factor receptor 1. Mol Cell Biol 2007; 27: 2590–2600.
Agarwal MK, Ruhul Amin AR, Agarwal ML . DNA replication licensing factor minichromosome maintenance deficient 5 rescues p53-mediated growth arrest. Cancer Res 2007; 67: 116–121.
Pandit B, Halasi M, Gartel AL . p53 negatively regulates expression of FoxM1. Cell Cycle 2009; 8: 3425–3427.
Chauhan BK, Reed NA, Zhang W, Duncan MK, Kilimann MW, Cvekl A . Identification of genes downstream of Pax6 in the mouse lens using cDNA microarrays. J Biol Chem 2002; 277: 11539–11548.
Oren M . Decision making by p53: life, death and cancer. Cell Death Differ 2003; 10: 431–442.
Wolf LV, Yang Y, Wang J, Xie Q, Braunger B, Tamm ER et al. Identification of pax6-dependent gene regulatory networks in the mouse lens. PLoS One 2009; 4: e4159.
Deshaies RJ, Joazeiro CA . RING domain E3 ubiquitin ligases. Annu Rev Biochem 2009; 78: 399–434.
Zhang G, Wang G, Wang S, Li Q, Ouyang G, Peng X . Applying proteomic methodologies to analyze the effect of hexamethylene bisacetamide (HMBA) on proliferation and differentiation of human gastric carcinoma BGC-823 cells. Int J Biochem Cell Biol 2004; 36: 1613–1623.
Dehay C, Kennedy H . Cell-cycle control and cortical development. Nat Rev Neurosci 2007; 8: 438–450.
Gotz M, Huttner WB . The cell biology of neurogenesis. Nat Rev Mol Cell Biol 2005; 6: 777–788.
Kuan CY, Roth KA, Flavell RA, Rakic P . Mechanisms of programmed cell death in the developing brain. Trends Neurosci 2000; 23: 291–297.
Green DR, Kroemer G . Cytoplasmic functions of the tumour suppressor p53. Nature 2009; 458: 1127–1130.
Suter DM, Cartier L, Bettiol E, Tirefort D, Jaconi ME, Dubois-Dauphin M et al. Rapid generation of stable transgenic embryonic stem cell lines using modular lentivectors. Stem Cells 2006; 24: 615–623.
Bonnefont J, Daulhac L, Etienne M, Chapuy E, Mallet C, Ouchchane L et al. Acetaminophen recruits spinal p42/p44 MAPKs and GH/IGF-1 receptors to produce analgesia via the serotonergic system. Mol Pharmacol 2007; 71: 407–415.
Livak KJ, Schmittgen TD . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25: 402–408.
Acknowledgements
JB was supported by the Auvergne Regional Council, the Geneva Department of Public Education and FP7-2007 ICT-216593 (SECO). CD was supported by FP7-2007 ICT-216593 (SECO) and ANR-06-NEUR-010. KHK was supported by the Swiss National Science Foundation.
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Bonnefont, J., Laforge, T., Plastre, O. et al. Primate-specific RFPL1 gene controls cell-cycle progression through cyclin B1/Cdc2 degradation. Cell Death Differ 18, 293–303 (2011). https://doi.org/10.1038/cdd.2010.102
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DOI: https://doi.org/10.1038/cdd.2010.102
