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Role of the Protein 14-3-3 in Spermatogenesis and Sperm Motility

Puri, Pawan
http://rave.ohiolink.edu/etdc/view?acc_num=kent1247848954

Abstract Details

2009, PHD, Kent State University, College of Arts and Sciences / Department of Biological Sciences.
The 14-3-3 proteins are a highly conserved family of acidic proteins and interact with various cellular phosphoproteins. The ability to bind phosphorylated proteins implicates 14-3-3 proteins in a variety of cell signaling pathways. While 14-3-3 protein isoforms are expressed in the testis and the sperm, their role in the spermatogenesis and the male gamete function is not known. The main goals of this dissertation are to identify the14-3-3 binding proteins in the sperm and the testis and to determine the physiological significance of the 14-3-3 binding interactions in the spermiogenesis and the sperm function. The sperm 14-3-3 binding proteins were isolated by GST-14-3-3-affinity chromatography. The 14-3-3 binding proteins in sperm can be classified as those involved in: fertilization, acrosome reaction, metabolism, protein folding and ubiquitin-mediated proteolysis. Identification of sperm PP1γ2 binding proteins by microcystin-agarose chromatography showed that PP1γ2 and 14 3-3 interactomes contain several common proteins. These proteins are likely to be phosphoproteins and potential PP1γ2 substrates. To purify 14-3-3 binding proteins in testis, transgenic mice expressing TAP-tag 14-3-3 were used. Tandem affinity purification (TAP) coupled with tandem mass spectrometry, identified more than one hundred 14-3-3 binding proteins in the testis. The 14-3-3 binding proteins in the testis included, proteins involved in spermatogenesis and variety of other cellular processes. The 14 3-3 interactome in the testis suggests an important regulatory role of the protein 14-3-3 in male germ cell divisions, differentiation and other cellular processes. To determine the physiological significance of the identified 14-3-3 interactions, transgenic mice expressing the 14-3-3 inhibitor peptide, difopein, in postmeiotic spermatids, were generated. Transgenic mice expressing the YFP-difopein have no apparent phenotype and have comparable fertility to their wild type littermates; hence difopein did not affect normal sperm function. Although this attractive approach was unable to determine the physiological significance of 14-3-3 interactions, the analysis suggests alternate strategies to answer this complex problem. In summary, the findings from this study add to the growing list of 14-3-3 interactors and these interactions suggest a role for 14-3-3 in integrating the regulation of signaling, metabolism and other cellular processes in testicular somatic cells, germ cells and mature spermatozoa.
Srinivasan Vijayaraghavan, PhD (Advisor)
Douglas Kline, PhD (Committee Member)
Jennifer Marcinkeiwicz, PhD (Committee Member)
Kenneth Rosenthal, PhD (Committee Member)
Soumitra Basu, PhD (Other)
149 p.
Biology
Sperm; 14-3-3; PP1; Difopein

Recommended Citations

Citations

  • Puri, P. (2009). Role of the Protein 14-3-3 in Spermatogenesis and Sperm Motility [Doctoral dissertation, Kent State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=kent1247848954

    APA Style (7th edition)

  • Puri, Pawan. Role of the Protein 14-3-3 in Spermatogenesis and Sperm Motility. 2009. Kent State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=kent1247848954.

    MLA Style (8th edition)

  • Puri, Pawan. "Role of the Protein 14-3-3 in Spermatogenesis and Sperm Motility." Doctoral dissertation, Kent State University, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=kent1247848954

    Chicago Manual of Style (17th edition)

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© 2009, all rights reserved.
This open access ETD is published by Kent State University and OhioLINK.