Submission Date

4-23-2024

Document Type

Paper- Restricted to Campus Access

Department

Biology

Adviser

Rebecca Lyczak

Committee Member

Rebecca Roberts

Committee Member

Karen Clemente

Department Chair

Dale Cameron

Project Description

The gonad of Caenorhabditis elegans houses the developing oocytes that will eventually pass through the spermatheca to become fertilized and begin the first embryonic cell division. This study focuses on a puromycin sensitive aminopeptidase, PAM-1, and its mutant effects of the development of oocytes in the gonad. It is known pam-1 mutants delay meiosis exit timing, and cause polarity issues that therefore end in low embryonic viability of the embryos. WEE-1.3 is an inhibitory kinase that reduces the activity of cyclin dependent kinase (CDK-1). WEE-1.3 interacts with PAM-1during oocyte maturation. Without WEE-1.3, precocious oocyte maturation occurs in the gonad, however, when pam-1 is mutated, this does not occur. Because of this interaction, we wanted to test if PAM-1 may also interact with CDK-1. This research focuses on the localization and activity of CDK-1 in C. elegans gonads in wild-type as compared to pam-1 mutants. To determine the presence of CDK-1 in the gonad, we used antibody staining and found significantly higher levels of CDK-1 in wildtype versus pam-1 mutants. Specifically, in the –1 oocyte position higher mean intensity of CDK-1 was present in the cytoplasm. While trying to determine the activity of CDK-1 with a sensor that localizes to the cytoplasm with activity, differences in chromosome alignment were noticed. In the presence of pam-1, oocyte positions –4 and –5 maintained random chromosomal alignment versus nuclear envelope alignment seen in those positions in the sensor strain. The mean intensity of the –1 oocyte is trending toward significance leading conclusions to be drawn that there may be less active CDK-1.

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