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Investigation of mlh-3 & mlh-1 in C. elegans

Bracho, Asier (2023) Investigation of mlh-3 & mlh-1 in C. elegans. Master's Thesis, University of Pittsburgh. (Unpublished)

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Abstract

Mlh1 and Mlh3 are genes found in mammals with roles in meiosis and DNA mismatch repair. Their protein products function as heterodimers that resolve double-Holliday junctions during meiosis I. In mammals, knockouts of both genes result in infertile mice, while in C. elegans, homozygous knockouts of mlh-1 appear viable. We identified the C. elegans gene ZK1098.3 as a possible Mlh3 homolog due to the similar predicted functions in their products. We hypothesize that ZK1098.3 is the C. elegans homolog of Mlh3 (mlh-3), and that it, together with mlh-1, may be functioning to repair non-canonical COs. We procured or produced knockouts of both genes and crossed them into meiotically sensitive backgrounds. We also tagged ZK1098.3 with V5. Our results showed localization of ZK1098.3 protein in the meiotic portion of the worm germ line, supporting the hypothesis that this gene is involved in meiosis. One of the mlh-1 KOs produced extra DAPI bodies in diakinesis and showed slow growth. However, the other mlh-1 KO, along with the mlh-3 KOs and various crosses into sensitized backgrounds, appeared phenotypically normal. This leads us to the conclusion that while mlh-1 and ZK1098.3 may have roles in meiosis, they are only minor, and further work is necessary to understand their functions.

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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Bracho, Asierasb208@pitt.eduasb2080000-0002-8863-4707
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Thesis AdvisorYanowitz, Judith L.yanowitzjl@mwri.magee.edu
Committee MemberRoman, Beth L.romanb@pitt.edu
Committee MemberPadiath, Quasarqpadiath@pitt.edu
Date: 9 May 2023
Date Type: Publication
Defense Date: 17 April 2023
Approval Date: 9 May 2023
Submission Date: 27 April 2023
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Number of Pages: 58
Institution: University of Pittsburgh
Schools and Programs: School of Public Health > Human Genetics
Degree: MS - Master of Science
Thesis Type: Master's Thesis
Refereed: Yes
Uncontrolled Keywords: C. elegans meiosis crossover mlh1 mlh3 mlh-1 mlh-3 nondisjunction germ line
Date Deposited: 10 May 2023 02:42
Last Modified: 10 May 2023 02:42
URI: http://d-scholarship.pitt.edu/id/eprint/44765

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