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Regulation of the desaturase

Project Full Title:

Regulation of the stearoyl-CoA desaturase

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Project Description:

This interdisciplinary project incorporates the disciplines of Biology and Chemistry . However the project is specifically housed within the Hope College Department of Biology. This project is only open to Hope College students.

Problems with the regulation of lipid metabolism contribute to many chronic human disorders including heart disease, diabetes, obesity, and even some cancers. The long-term goal of the work is to gain a better understanding of the regulation of lipid production by dietary fats. We focus on one enzyme, the stearoyl-CoA fatty acid desaturase, which is encoded by the OLE1 gene in the model organism Saccharomyces cerevisiae. The desaturase produces monounsaturated fatty acids from saturated precursors. It is strongly regulated by the availability of fats in the diet. While aspects of the regulation are understood, it is far from complete: cells regulate expression of OLE1 through the ER resident transcription factors Mga2p and Spt23p. These proteins are switched on from an inactive p120 form to an active p90 form, which translocate into the nuclease and activate transcription of OLE1. What signal causes Mga2p and Spt23p to be switched from “off” to “on”? What proteins are involved in these processes? How do cells sense the type and presence of fed fatty acids? The overarching hypothesis of this work is that fed fatty acids are trafficked to internal membranes where protein sensors recognize and communicate the status of the membrane to these regulators of OLE1 gene expression. The objective of the work this summer will be to identify the signals and gene products that regulate expression of OLE1, and how they work.

Students working on this project will use both molecular genetic and biochemical approaches in their work. Experimental procedures will include some or all of the following: cell culture, cloning, DNA isolation, PCR, qPCR, gel electrophoresis, spectrophotometry, reporter gene assays, protein-protein interactions using 2 hybrid analysis, western blotting, GC and GC-MS, and microscopy. In addition to experimentation, students will be expected to be full members of the research team-analyzing data, preparing figures, reading and discussing research literature, and to present their results at a scientific conference.

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