Piwi/Argonaute (Ago) is the only known protein family with function in stem cell self-renewal highly conserved in both animal and plant kingdoms. These proteins also play key roles in germline development and RNAi/miRNA-mediated mechanisms. Our long term goal is to learn molecular mechanisms mediated by the Piwi/Ago proteins in regulating spermatogenesis. This will also contribute to understanding of stem cell division and other developmental processes mediated by this family of proteins. Our strategy has been to use Drosophila as a model to study these proteins in gametogenesis, and then to further what we learn from Drosophila to mammalian and clinical settings. Particularly, in the current funding period, we have demonstrated that mili, a murine member of the gene family, is essential for spermatogonial stem cell division and meiosis, yet miwi, another member, is a key regulator of spermiogenesis. These represent the first two in vivo studies of the piwi/ago gene family in mammalian systems. Moreover, we have shown that the overexpression of a human piwi gene, hiwi, is highly correlated to testicular seminomas.
Our specific aim here is to study the molecular mechanisms through which mili and miwi regulate spermatogenesis. Piwi/Ago family proteins are known to negatively regulate gene expression via RNAi, miRNA, and epigenetic pathways. Our latest studies, however, indicate that miwi and mili positively regulate translation and stability of target mRNAs, possibly via a miRNA-mediated mechanism. These findings provide an unparalleled opportunity for us to discover a novel mechanism of gene regulation that is fundamental to reproductive and stem cell biology. Our working hypothesis has three components: First, Mili and Miwi bind to and stabilize target mRNAs required for their target spermatogenic events;second, this function is coupled to translational regulation;third, this function is mediated by miRNAs. To test this hypothesis, we propose to: (1) systematically identify target mRNAs of Mili and Miwi. (2) Characterize the effect of Mili and Miwi binding on the stability of target mRNAs. (3) Determine if Mili and Miwi controls the stability of target mRNAs by regulating their translation. (4) Identify Mili/Miwi-target miRNAs and determine their function in regulating mRNA. Because Piwi/Ago proteins have demonstrated roles in oncogenesis, the proposed studies should bear significant health implications.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD042012-09
Application #
7795766
Study Section
Cellular, Molecular and Integrative Reproduction Study Section (CMIR)
Program Officer
Moss, Stuart B
Project Start
2002-04-01
Project End
2012-02-28
Budget Start
2010-03-01
Budget End
2011-02-28
Support Year
9
Fiscal Year
2010
Total Cost
$321,136
Indirect Cost
Name
Yale University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Ge, Xin Quan; Lin, Haifan (2014) Noncoding RNAs in the regulation of DNA replication. Trends Biochem Sci 39:341-3
Ross, Robert J; Weiner, Molly M; Lin, Haifan (2014) PIWI proteins and PIWI-interacting RNAs in the soma. Nature 505:353-359
Lin, Haifan (2014) Special Topic: Frontiers in RNA Research: The ever-expanding RNA world. Natl Sci Rev 1:182
Watanabe, Toshiaki; Lin, Haifan (2014) Posttranscriptional regulation of gene expression by Piwi proteins and piRNAs. Mol Cell 56:18-27
Cheng, Ee-chun; Lin, Haifan (2013) Repressing the repressor: a lincRNA as a MicroRNA sponge in embryonic stem cell self-renewal. Dev Cell 25:1-2
Nolde, Mona J; Cheng, Ee-Chun; Guo, Shangqin et al. (2013) Piwi genes are dispensable for normal hematopoiesis in mice. PLoS One 8:e71950
Saxe, Jonathan P; Chen, Mengjie; Zhao, Hongyu et al. (2013) Tdrkh is essential for spermatogenesis and participates in primary piRNA biogenesis in the germline. EMBO J 32:1869-85
Lin, Haifan (2012) Capturing the cloud: UAP56 in nuage assembly and function. Cell 151:699-701
Chen, Dong; Zheng, Wei; Lin, Aiping et al. (2012) Pumilio 1 suppresses multiple activators of p53 to safeguard spermatogenesis. Curr Biol 22:420-5
Wang, Jianguan; Gu, Honggang; Lin, Haifan et al. (2012) Essential roles of the chromatin remodeling factor BRG1 in spermatogenesis in mice. Biol Reprod 86:186

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