Germ cells carry out the reproductive function of multicellular organisms, and normal germ cell development ensures survival of the species. Germ granules are conserved cytoplasmic organelles of germ cells, essential for survival, differentiation, and function of these cells. Mutations in germ granule components or loss of their expression lead to infertility in model organisms and are associated with infertility in humans. By contrast, inappropriate expression of germ granule components in somatic cells is linked to carcinogenesis in humans. Many RNA-binding proteins and developmentally regulated mRNAs are found enriched in germ granules, leading to a hypothesis that these organelles function in regulation of mRNA stability or translational activity; yet, the molecular function of these organelles is still undefined. The nematode C. elegans has been instrumental for analysis of mRNA translational control in germline development. Through focusing on cofactors that promote localization of RNA-binding regulatory protein FBF-2 to germ granules (called P granules in this species) in germline stem cells we have generated a set of innovative tools that will address key mechanisms through which P granules regulate FBF-2 activity. By integrating genetic, molecular, biochemical and imaging-based approaches, we will: 1) Determine how FBF-2 interaction with a cofactor protein DLC-1 important for P granule localization impacts translational repression exerted by FBF-2; 2) Define the mechanisms of P granule remodeling that specifically degrade FBF-2 at the onset of meiosis when stem cells transition from proliferation to differentiation; 3) Determine the role of DLC-1 binding to core P granule components in regulation of stability of embryonic P granules and in recruitment of transient P granule components such as FBF-2 to adult germline P granules. Our experimental system is poised to reveal specific molecular mechanisms mediating the interplay between germ granules and translational regulation. Since germ granules are conserved organelles and our research focuses on conserved regulatory proteins, studies in this model system will provide critical insight into the causes of infertility in humans.

Public Health Relevance

Germ granules are conserved organelles essential for animal reproduction as they support development and function of germ cells. Using the power of genetic analysis available in C. elegans, studies of conserved regulatory proteins will uncover the mechanisms mediating the interplay between germ granules and post- transcriptional control in human germ cells. In the long term, this work will help understanding germ cell dysfunction leading to human infertility or misregulation of germ-cell-specific regulatory pathways leading to carcinogenesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM109053-06
Application #
9739573
Study Section
Cellular, Molecular and Integrative Reproduction Study Section (CMIR)
Program Officer
Salazar, Desiree Lynn
Project Start
2014-03-01
Project End
2023-05-31
Budget Start
2019-09-01
Budget End
2020-05-31
Support Year
6
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of Montana
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
010379790
City
Missoula
State
MT
Country
United States
Zip Code
59812
Day, Nicholas J; Ellenbecker, Mary; Voronina, Ekaterina (2018) Caenorhabditis elegans DLC-1 associates with ribonucleoprotein complexes to promote mRNA regulation. FEBS Lett 592:3683-3695
Ellenbecker, Mary; Osterli, Emily; Wang, Xiaobo et al. (2018) Dynein Light Chain DLC-1 Facilitates the Function of the Germline Cell Fate Regulator GLD-1 in Caenorhabditis elegans. Genetics :
Battisti, James M; Watson, Lance A; Naung, Myo T et al. (2017) Analysis of the Caenorhabditis elegans innate immune response to Coxiella burnetii. Innate Immun 23:111-127
Voronina, Ekaterina (2016) mRNAs Hit a Sticky Wicket. Dev Cell 37:9-10
Su, Zhangli; Wang, Fengbin; Lee, Jin-Hee et al. (2016) Reader domain specificity and lysine demethylase-4 family function. Nat Commun 7:13387
Wang, Xiaobo; Olson, Jenessa R; Rasoloson, Dominique et al. (2016) Dynein light chain DLC-1 promotes localization and function of the PUF protein FBF-2 in germline progenitor cells. Development 143:4643-4653
Novak, Preston; Wang, Xiaobo; Ellenbecker, Mary et al. (2015) Splicing Machinery Facilitates Post-Transcriptional Regulation by FBFs and Other RNA-Binding Proteins in Caenorhabditis elegans Germline. G3 (Bethesda) 5:2051-9