Date

2018

Department or Program

Biological Chemistry

Primary Wellesley Thesis Advisor

John Goss

Additional Advisor(s)

Kaye Peterman

Additional Advisor

Donald Elmore

Additional Advisor

Markella Rutherford

Abstract

The stages of cellular division are regulated by highly conserved proteins, many of which are essential for cell viability. Modifying the function of essential proteins through traditional genetic approaches is limited because deleting or altering essential genes often results in cell death. Since the subcellular localization of a protein contributes to its cellular function, transiently altering protein localization can temporarily disrupt function and provide insight into how a protein of interest contributes to a cellular pathway in real time. Light-inducible dimerization modules utilized in optogenetics are tools that can enable rapid and reversible spatio-temporal manipulation of in vivo protein localization, thereby achieving transient disruption of function.

In this study, we utilized molecular cloning to design an optogenetic, blue light-inducible dimerization system using genetically encoded CRY2 and CIBN plant cryptochromes. The CRY2/CIBN cryptochrome module has been previously implemented in mammalian systems to manipulate cellular processes such as phosphoinositide metabolism, circadian rhythm, and gene transcription. Our genetically encoded CRY2 and CIBN cryptochrome fusion tags can be used to alter the spatio-temporal localization of any protein of interest in the fission yeast, Schizosaccharomyces pombe. Fission yeast are a valuable model organism for studying regulation of cell division due to facile genetic manipulation, compatibility with microscopy, and high levels of conservation with eukaryotic genes. This novel approach to studying fission yeast cytokinesis will enable us to better investigate the role essential proteins play in regulating processes during cellular division.

Available for download on Tuesday, May 23, 2023

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