GSBS Dissertations and Theses

ORCID ID

0000-0001-7643-4516

Publication Date

2018-07-10

Document Type

Doctoral Dissertation

Academic Program

Interdisciplinary Graduate Program

Department

RNA Therapeutics Institute

First Thesis Advisor

Craig Mello

Keywords

CRISPR-Cas9-induced indels; CRISPR-Cas9-mediated HR; Co-CRISPR, PIE-1, SUMOylation, C. elegans, germline specification, germline integrity, NuRD complex, HDA-1

Abstract

In many organisms, the most fundamental event during embryogenesis is differentiating between germline cells and specialized somatic cells. In C. elegans, PIE-1 functions to protect the germline from somatic differentiation and appears to do so by blocking transcription and by preventing chromatin remodeling in the germline during early embryogenesis. Yet the molecular mechanisms by which PIE-1 specifies germline remain poorly understood. Our work shows that SUMOylation facilitates PIE-1-dependent germline maintenance and specification. In vivo SUMO purification in various CRISPR strains revealed that PIE-1 is SUMOylated at lysine 68 in the germline and that this SUMOylation is essential for forming NuRD complex and preserving HDA-1 activity. Moreover, HDA-1 SUMOylation is dependent on PIE-1 and enhanced by PIE-1 SUMOylation, which is required for protecting germline integrity. Our results suggest the importance of SUMOylation in the germline maintenance and exemplify simultaneous SUMOylation of proteins in the same functional pathway.

DOI

10.13028/verx-az15

Rights and Permissions

Copyright is held by the author, with all rights reserved.

Available for download on Saturday, August 24, 2019

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