One major goal of this proposal is to determine the molecular mechanism of temporal translational control during male germ cell development in the mouse. A second major objective is to investigate the role of the protamine 1 gene in the physical remodelling and the genetic reprogramming of the paternal genome during spermatogenesis. The mouse protamine 1 gene, Prm-1, is regulated at both the transcriptional and translational level. Although the gene is first transcribed postmeiotically in round spermatids, the mRNA is not translated until up to a week later in elongating spermatids. To identify the cis-acting regulatory elements in the Prm-1 mRNA that regulate its temporal translational control, fusions will be constructed between regions of the Prm-1 gene and a reporter gene and studied in transgenic mice. Cis-acting translational control elements that are identified will be cloned into different regions of an heterologous mRNA to determine if their position within an mRNA affects their function. To study the mechanism of translational control, genetic and biochemical approaches will be used to clone the gene(s) for translational control factor(s) that interact with the translational control elements. To determine the importance of temporal translational control as it pertains to protamine 1 function, the product of the Prm-1 gene will be expressed at different times during spermatogenesis. To determine if protamine 1 is essential for nuclear condensation and/or reprogramming of the paternal genome during spermatogenesis, the gene will be mutated in embryonic stem cells by homologous recombination and its effect studied in transgenic mice. Knowledge gained from these studies may ultimately contribute to the medically important problems of male infertility and the inheritance of genetic diseases.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD027215-02
Application #
3328813
Study Section
Reproductive Biology Study Section (REB)
Project Start
1990-09-01
Project End
1995-04-30
Budget Start
1991-05-01
Budget End
1992-04-30
Support Year
2
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Washington
Department
Type
Schools of Arts and Sciences
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195
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Zhong, J; Peters, A H; Kafer, K et al. (2001) A highly conserved sequence essential for translational repression of the protamine 1 messenger rna in murine spermatids. Biol Reprod 64:1784-9
Giorgini, F; Davies, H G; Braun, R E (2001) MSY2 and MSY4 bind a conserved sequence in the 3' untranslated region of protamine 1 mRNA in vitro and in vivo. Mol Cell Biol 21:7010-9
Nadler, J J; Braun, R E (2000) Fanconi anemia complementation group C is required for proliferation of murine primordial germ cells. Genesis 27:117-23

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