Abstract

The long-term goal of these studies is to determine how synthesis of proteins is regulated post-transcriptionally during spermatogenesis. The first specific aim is to purify and characterize the protein. Here the principal investigator proposes to take advantage of RNA affinity chromatography methods, where the protein will bind to its target RNA sequences immobilized on sepharose. Purified protein will be analyzed by microsequencing.
The second aim will be to isolate cDNA for TB-RBP as well as construct antibody and to use these reagents to define the extent of the TB-RBP gene families and to determine the cellular and subcellular distribution of TB-RBP.
The third aim i s to determine the RNA recognition sites of the TB-RBP. This will be done by methods of site-directed mutagenesis of the target RNA sequences and the end goal is to better understand how TB-RBP binds to and inhibits mRNA translation.
The fourth aim i s to define the sites and mechanisms of phosphorylation and dephosphorylation of TB-RBP.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD028832-20
Application #
2889051
Study Section
Special Emphasis Panel (ZRG2-END (02))
Program Officer
Tasca, Richard J
Project Start
1980-09-01
Project End
2001-08-31
Budget Start
1999-09-01
Budget End
2000-08-31
Support Year
20
Fiscal Year
1999
Total Cost
Indirect Cost

  Institution

Name
University of Pennsylvania
Department
Obstetrics & Gynecology
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104

  Publications

Xu, Mingang; McCarrey, John R; Hecht, Norman B (2008) A cytoplasmic variant of the KH-type splicing regulatory protein serves as a decay-promoting factor for phosphoglycerate kinase 2 mRNA in murine male germ cells. Nucleic Acids Res 36:7157-67
Yu, Zuoren; Hecht, Norman B (2008) The DNA/RNA-binding protein, translin, binds microRNA122a and increases its in vivo stability. J Androl 29:572-9
Xu, Mingang; Hecht, Norman B (2008) MSY2 and polypyrimidine tract binding protein 2 stabilize mRNAs in the mammalian testis. Int J Androl 31:457-61
Xu, Mingang; You, Yun; Hunsicker, Patricia et al. (2008) Mice deficient for a small cluster of Piwi-interacting RNAs implicate Piwi-interacting RNAs in transposon control. Biol Reprod 79:51-7
Iguchi, Naoko; Xu, Mingang; Hori, Tamaki et al. (2007) Noncoding RNAs of the mammalian testis: the meiotic transcripts Nct1 and Nct2 encode piRNAs. Ann N Y Acad Sci 1120:84-94
Xu, Mingang; Hecht, Norman B (2007) Polypyrimidine tract binding protein 2 stabilizes phosphoglycerate kinase 2 mRNA in murine male germ cells by binding to its 3'UTR. Biol Reprod 76:1025-33
Stein, Joel M; Bergman, Wayland; Fang, Yanshan et al. (2006) Behavioral and neurochemical alterations in mice lacking the RNA-binding protein translin. J Neurosci 26:2184-96
Iguchi, Naoko; Tobias, John W; Hecht, Norman B (2006) Expression profiling reveals meiotic male germ cell mRNAs that are translationally up- and down-regulated. Proc Natl Acad Sci U S A 103:7712-7
Yu, Zuoren; Raabe, Tobias; Hecht, Norman B (2005) MicroRNA Mirn122a reduces expression of the posttranscriptionally regulated germ cell transition protein 2 (Tnp2) messenger RNA (mRNA) by mRNA cleavage. Biol Reprod 73:427-33
Cho, Yoon Shin; Iguchi, Naoko; Yang, Juxiang et al. (2005) Meiotic messenger RNA and noncoding RNA targets of the RNA-binding protein Translin (TSN) in mouse testis. Biol Reprod 73:840-7

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