Abstract

The long-term objective of our research is to understand the biological function of a family of transaltional modulators known as PABP interacting protiens (Paips) which were first cloned in our laboratory. These proteins mediate their effects on translation by interacting with the poly(A) binding protein (PABP). Previous work has shown that Paipi stimulates translation, whereas Paip2A and Paip2B inhibit translation.
The specific aims of this proposal are to further our understanding of the molecular mechanism of Paip action, to discover how Paip function is regulated, and to elucidate the biological significance of this family of proteins. A variety of in vitro and in vivo experiments will be carried out to study the Paip1PABP interactions, to dissect their mechanism of action, and to identify novel Paip interacting proteins. Paip expression and function will be inhibited by the RNA interference technique and the use of small cell-permeable peptides. Paip phosphorylation will be explored to elucidate the signaling pathways impinging upon Paip function. To this end, phosphopeptide mapping and 2-dimensional isoelectric focusing and SDS-PAGE will be used to study the phosphorylation states of the Paips under various environmental conditions and upon treatment with pharmacological kinase inhibitors. The phosphoresidues in the proteins will be identified and mutated. Resulting proteins will be assayed using in vitro and in vivo translation experiments to discover their functional importance. Elucidation of the physiological role of the Paips will be pursued by Paip overexpression and knockout (KO) experiments in Drosophila. Paip null flies will be generated by P-element insertion or homologous recombination. The generation of KO mice devoid of the individual Paip genes will also be pursued by homologous recombination. In the case of Paip2A and Paip2B, which are functional homologs, a Paip2AIPaip2B KO mouse will be generated. KO mice will be analyzed for phenotypic abnormalities.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM066157-03
Application #
6791292
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Rhoades, Marcus M
Project Start
2002-09-01
Project End
2006-08-31
Budget Start
2004-09-01
Budget End
2005-08-31
Support Year
3
Fiscal Year
2004
Total Cost
$189,000
Indirect Cost

  Institution

Name
Mcgill University
Department
Type
DUNS #
205667090
City
Montreal
State
PQ
Country
Canada
Zip Code
H3 0-G4

  Publications

Cao, Ruifeng; Gkogkas, Christos G; de Zavalia, Nuria et al. (2015) Light-regulated translational control of circadian behavior by eIF4E phosphorylation. Nat Neurosci 18:855-62
Cao, Ruifeng; Robinson, Barry; Xu, Haiyan et al. (2013) Translational control of entrainment and synchrony of the suprachiasmatic circadian clock by mTOR/4E-BP1 signaling. Neuron 79:712-24
Khoutorsky, Arkady; Yanagiya, Akiko; Gkogkas, Christos G et al. (2013) Control of synaptic plasticity and memory via suppression of poly(A)-binding protein. Neuron 78:298-311
Svitkin, Yuri V; Yanagiya, Akiko; Karetnikov, Alexey E et al. (2013) Control of translation and miRNA-dependent repression by a novel poly(A) binding protein, hnRNP-Q. PLoS Biol 11:e1001564
Rosenfeld, Amy B (2011) Suppression of cellular transformation by poly (A) binding protein interacting protein 2 (Paip2). PLoS One 6:e25116
Alvarez-Saavedra, Matías; Antoun, Ghadi; Yanagiya, Akiko et al. (2011) miRNA-132 orchestrates chromatin remodeling and translational control of the circadian clock. Hum Mol Genet 20:731-51
Bidinosti, Michael; Ran, Israeli; Sanchez-Carbente, Maria R et al. (2010) Postnatal deamidation of 4E-BP2 in brain enhances its association with raptor and alters kinetics of excitatory synaptic transmission. Mol Cell 37:797-808
Yanagiya, Akiko; Delbes, Geraldine; Svitkin, Yuri V et al. (2010) The poly(A)-binding protein partner Paip2a controls translation during late spermiogenesis in mice. J Clin Invest 120:3389-400
Svitkin, Yuri V; Evdokimova, Valentina M; Brasey, Ann et al. (2009) General RNA-binding proteins have a function in poly(A)-binding protein-dependent translation. EMBO J 28:58-68
Kanaan, Ahmad Seif; Frank, Filipp; Maedler-Kron, Chelsea et al. (2009) Crystallization and preliminary X-ray diffraction analysis of the middle domain of Paip1. Acta Crystallogr Sect F Struct Biol Cryst Commun 65:1060-4

Showing the most recent 10 out of 21 publications