The B-regulatory subunit of protein phosphatase 2A mediates the dephosphorylation of rice retinoblastoma-related protein-1

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• 作者：Edit ábrahám (1)
  Ping Yu (1)
  Ilona Farkas (2)
  Zsuzsanna Darula (3)
  Erzsébet Varga (4)
  Noémi Lukács (4)
  Ferhan Ayaydin (5)
  Katalin F. Medzihradszky (3)
  Viktor Dombrádi (2)
  Dénes Dudits (1)
  Gábor V. Horváth (1)
  
• 关键词：Cell cycle regulation ; Retinoblastoma ; related protein ; Protein phosphatase 2A ; Cyclin ; dependent protein kinase ; Protein phosphorylation
• 刊名：Plant Molecular Biology
• 出版年：2015
• 出版时间：January 2015
• 年：2015
• 卷：87
• 期：1-2
• 页码：125-141
• 全文大小：1,250 KB
• 参考文献：1. ábrahám E, Miskolczi P, Ayaydin F, Yu P, Kotogány E, Bakó L, ?tv?s K, Horváth GV, Dudits D (2011) Immunodetection of retinoblastoma-related protein and its phosphorylated form in interphase and mitotic alfalfa cells. J Exp Bot 62:2155-168. doi:10.1093/jxb/erq413 CrossRef
  2. Ach RA, Durfee T, Miller AB, Taranto P, Hanley-Bowdoin L, Zambryski PC, Gruissem W (1997) RRB1 and RRB2 encode maize retinoblastoma-related proteins that interact with a plant D-type cyclin and geminivirus replication protein. Mol Cell Biol 17:5077-086
  3. Avni D, Yang H, Martelli F, Hofmann F, ElShamy WM, Ganesan S, Scully R, Livingston DM (2003) Active localization of the retinoblastoma protein in chromatin and its response to S phase DNA damage. Mol Cell 12:735-46 CrossRef
  4. Ayaydin F, Kotogány E, Abrahám E, Horváth GV (2011) Synchronization of / Medicago sativa cell suspension culture. Methods Mol Biol 761:227-38. doi:10.1007/978-1-61779-182-6_15 CrossRef
  5. Barr FA, Elliott PR, Gruneberg U (2011) Protein phosphatases and the regulation of mitosis. J Cell Sci 124:2323-334. doi:10.1242/jcs.087106 CrossRef
  6. Berndt N, Dohadwala M, Liu CWY (1997) Constitutively active protein phosphatase 1 alpha causes Rb dependent G1 arrest in human cancer cells. Curr Biol 7:375-86 CrossRef
  7. Bianchi M, Villa-Moruzzi E (2001) Binding of phosphatase-1 delta to the retinoblastoma protein pRb involves domains that include substrate recognition residues and a pRB binding motif. Biochem Biophys Res Commun 280:1- CrossRef
  8. Biemann K (1990) Appendix 5. Nomenclature for peptide fragment ions (positive ions). Methods Enzymol 193:886-87 CrossRef
  9. Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513-523 CrossRef
  10. Bollen M, Peti W, Ragusa MJ, Beullens M (2010) The extended PP1 toolkit: designed to create specificity. Trends Biochem Sci 35:450-58. doi:10.1016/j.tibs.2010.03.002 CrossRef
  11. Boniotti MB, Gutierrez C (2001) A cell-cycle-regulated kinase activity phosphorylates plant retinoblastoma protein and contains, in Arabidopsis, a CDKA/cyclin D complex. Plant J 28:341-50 CrossRef
  12. Boruc J, Mylle E, Duda M, De Clercq R, Rombauts S, Geelen D, Hilson P, Inzé D, Van Damme D, Russinova E (2010) Systematic localization of the Arabidopsis core cell cycle proteins reveals novel cell division complexes. Plant Physiol 152:553-65. doi:10.1104/pp.109.148643 CrossRef
  13. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248-54 CrossRef
  14. Cicchillitti L, Fasanaro P, Biglioli P, Capogrossi MC, Martelli F (2003) Oxidative stress induces protein phosphatase 2A-dependent dephosphorylation of the pocket proteins pRb, p107, and p130. J Biol Chem 278:19509-9517 CrossRef
  15. Cohen PT (2002) Protein phosphatase 1—targeted in many directions. J Cell Sci 115:241-56
  16. Davis AJ, Yan Z, Martinez B, Mumby MC (2008) Protein phosphatase 2A is targeted to cell division control protein 6 by a calcium-binding regulatory subunit. J Biol Chem 283:16104-6114
• 作者单位：Edit ábrahám (1)
  Ping Yu (1)
  Ilona Farkas (2)
  Zsuzsanna Darula (3)
  Erzsébet Varga (4)
  Noémi Lukács (4)
  Ferhan Ayaydin (5)
  Katalin F. Medzihradszky (3)
  Viktor Dombrádi (2)
  Dénes Dudits (1)
  Gábor V. Horváth (1)
  
  1. Institute of Plant Biology, Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary
  2. Department of Medical Chemistry, University of Debrecen, Debrecen, Hungary
  3. Laboratory of Proteomics Research, Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary
  4. Department of Plant Biology and Plant Biochemistry, Faculty of Horticultural Science, Corvinus University of Budapest, Budapest, Hungary
  5. Laboratory of Cellular Imaging, Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary
  
• ISSN：1573-5028

文摘

The phosphorylation of plant retinoblastoma-related (RBR) proteins by cyclin-dependent kinases (CDKs) is well documented, but the counteracting phosphatases have not been identified yet. We report here that rice retinoblastoma-related protein-1 (OsRBR1) interacted with the B-subunit of rice protein phosphatase 2A (OsPP2A B- and underwent reversible phosphorylation during the cell division cycle. The OsRBR1-OsPP2A B-association required B domain in OsRBR1 and the C-terminal region of OsPP2A B- We found by immunoprecipitation that OsPP2A B- OsPP2A catalytic subunit subtype II, PSTAIRE-type CDK and OsRBR1 were in the same protein complex, indicating a physical association between the phosphatase, the kinase and their common substrate. OsPP2A B-contains three predicted CDK phosphorylation sites: Ser95, Ser102 and Ser119. The in vitro phosphorylation of Ser95 and Ser119 with PSTAIRE-kinases was verified by mass spectrometry. We generated a series of phosphorylation site mutants to mimic the dephosphorylated or phosphorylated states of OsPP2A B- and confirmed that all of the three predicted sites can be phosphorylated. Yeast two-hybrid experiments suggested that the phosphorylation of OsPP2A B-promoted the formation of the OsPP2A holoenzyme. A triple phosphorylation mimicking OsPP2A B-mutant containing holoenzyme showed higher activity in phosphatase assays. Our data collectively show that the phosphatase activity of OsPP2A against OsRBR1 is regulated by the phosphorylation of its B-regulatory subunit. However, the analysis of the effect of okadaic acid, a phosphatase inhibitor, in rice cell suspension cultures revealed that the dephosphorylation of OsRBR1 was completely inhibited only by high dose (300?nM) of the okadaic acid during the cell cycle progression. Therefore the role of the protein phosphatase 1 should be considered as an additional post translational regulatory component of RBR protein function in higher plants.