Document Type

Thesis

Date of Award

5-2014

School/College

College of Science, Engineering, and Technology (COSET)

Degree Name

MS in Biology

First Advisor

Assistant Professor Shodimu-Emmanuel Olufemi

Abstract

Kinases and phosphatases regulate protein phosphorylation and dephosphorylation that playa major role in gene expression. One of the phosphatases, serine/threonine protein phosphatase 6 catalytic subunit (PPP6C), regulates the progression of the cell cycle by dephosphorylating (TAK1) transforming-growth factor 􁪽 activated 1 protein kinase. TAK1 is involved in controlling cellular functions and regulating transcription. The functions of PPP6C have been elucidated and proteins that regulate PPP6C have been identified and studied, but other regulators of PPP6C have not been fully studied. To identify other regulators of PPP6C and to characterize their functions, we focused on microRNAs (miRNAs) and identified in the miRBase database, miRNAs that regulate PPP6C mRNA; and we select one of the several miRNAs, hsamiR- 765, that regulates mRNA of PPP6C to inhibit its translation and cleave its mRNA. To decipher if hsa-miR-765 target mRNA of PPP6C, we cloned the stem-loop of hsamiR- 765 into pmR-ZsGreen1 expression vector, a green fluorescence protein (GFP) and express hsa-miR-765-GFP construct in PC-3 cells. We showed for the first time that over-expression of hsa-miR-765 targets mRNA of PPP6C in PC3 cells show a down 1 regulation of PPP6C expression in 24 hour intervals. It is likely that the downregulation of PPP6C protein causes an increase in TAKI, a target of PPP6C dephosphorylation. Our results validate the computational prediction information at the miRBase database that hsa-rriiR-765 target PPP6C mRNA.

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