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A Screen for Genetic Modifiers of Protein Phosphatase 1 in Drosophila Collective Cell Migration

Undergraduate #51
Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology

Carmen F. Del Real - Dodge City Community College
Co-Author(s): Jocelyn McDonald, Kansas State University, Manhattan, KS; Yujun Chen, Kansas State University, Manhattan, KS



Inside the body, some cells migrate collectively in tightly or loosely-associated groups. Cells migrate to shape tissues that form organs, for embryonic development, immune system responses, and in tumor metastases. Drosophila melanogaster provides an excellent genetic model to understand how groups of cells collectively migrate. Not only do their small, transparent tissues allow observations of cell migration but also, many human disease genes are conserved in Drosophila. During development of the ovary, 6-8 cells form the border cell cluster, which stay together during their entire migration to the large oocyte. We recently found that the Nuclear inhibitor of protein Ser/Thr phosphatase 1 (NiPP1) protein causes border cells to separate into single cells, become more rounded, and migrate much slower than normal. Therefore, overexpressing NiPP1 allows us, through a genetic modifier screen, to determine the Protein Phosphatase 1 (PP1) regulatory and catalytic subunits required for adhesion and collective migration of border cells. In addition, this type of screen can identify genes important for the correct function of PP1 in border cells. To carry out this screen, we crossed females expressing NiPP1 in border cells to a collection of mutant strains that remove large numbers of genes (called ‘deficiencies’). Ovaries from females of the F1 generation with the correct genotype were dissected and stained with antibodies to visualize, by fluorescent microscopy, the ability of border cells to migrate and stay together. More than 35 mutants were crossed to NiPP1, with several showing enhanced migration and one exhibiting better migration and cohesion of the border cell cluster. These mutants represent candidate PP1 regulatory genes. Future work includes performing further genetic screens using smaller deficiency mutant strains to determine the genes that modify NiPP1 phenotypes. Finally, we plan to use RNAi to knock down candidate genes one by one to definitively determine the genes required for cell-to-cell adhesion and collective migration. Because many Drosophila genes have human homologs, these studies on PP1 have implications for collective cell migration in human development and tumor metastasis.

Not Submitted

Funder Acknowledgement(s): Research is supported by National Science Foundation (NSF), Kansas Louis Stokes Alliance for Minority Participation under grant #1305059 and under NSF grant #1456053 to Jocelyn A. McDonald.

Faculty Advisor: Jocelyn A. McDonald, jmcdona@ksu.edu

Role: I worked with the post doc, Yujun Chen with this project under Jocelyn McDonalds' Lab. I was able to complete all 35 lines, however, Dr. Chen has completed over 150+ lines. The information provided in this abstract was my research, Yujun's research results was not provided.

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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