The goal of this project is to identify and characterize new germ plasm components that are required for primordial germ cell development in the nematode, Caenorhabditis elegans and, in so doing, gain a better understanding of the molecular mechanisms controlling germ cell development. To this end, two approaches will be used, one biochemical and one genetic, both of which will employ the germ plasm associated protein, PIE-1.
Specific Aim I, will use a biochemical approach to identify mRNAs that associate with PIE-1 in vivo. Endogenous PIE- 1 protein will be immunoprecipitated (IPed) from embryo protein extracts. mRNAs that bind to PIE-1 should co-IP, and can then be isolated by phenol/chloroform extraction.
In Specific Aim II, RNAi, in situ hybridization and immunofluorescent cytochemistry will be used to begin analyzing the function and possible role of the PIE- l associating RNAs in germ cell development.
Specific Aim III, will employ a genetic screen for maternal-effect-sterile mutants, performed in a pie-1/+ background. It is anticipated that this sensitized genetic background will allow the identification of previously undetectable maternal-effect-sterile phenotypes, specifically mutations that affect primordial germ cell development, and/or perhaps additional PIE-1 signaling elements.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM064201-03
Application #
6636703
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Tompkins, Laurie
Project Start
2001-07-01
Project End
2003-12-31
Budget Start
2003-07-01
Budget End
2003-12-31
Support Year
3
Fiscal Year
2003
Total Cost
$24,074
Indirect Cost
Name
Johns Hopkins University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218