{"@context":"http://iiif.io/api/presentation/2/context.json","@type":"sc:Manifest","@id":"https://iiif.quartexcollections.com/wesleyan/iiif/65cfc942-d3ea-48af-9a37-6109b489a7e8/manifest","label":"m2017-k97","metadata":[{"label":"Title","value":"Investigating the intersection between DNA mismatch repair and Triplet nucleotide repeat expansion"},{"label":"Subject","value":["Molecular biology","Trinucleotide repeats","DNA repair"]},{"label":"Rights","value":["In Copyright"]},{"label":"Repository","value":["University Archives"]},{"label":"Language","value":["English"]},{"label":"Item Information","value":"<a href=\"https://onesearch.wesleyan.edu/permalink/01CTW_WU/12cbh08/alma9932185924203768\" target=\"_blank\">M2017 K97</a>"},{"label":"Held By","value":"Wesleyan University Library, Wesleyan University, Middletown, CT, USA"},{"label":"Group","value":["Theses & Dissertations"]},{"label":"Name(s)","value":["Kutlu, Selin (creator)"]},{"label":"Administrative Note","value":"Node ID: 16930<br>Administrative Notes: Old URL: https://wesscholar.wesleyan.edu/etd_mas_theses/161<br>Author Contact: skutlu@wesleyan.edu<br>Local Identifier: 161_kutlu_s_thesis<br>PID: ir:2475"},{"label":"Author-supplied Keywords","value":["Kinetics","TNR expansion","Huntington's disease","Msh2-Msh3","Msh2-Msh6","Triplet nucleotide repeats","DNA mismatch repair"]},{"label":"Degree","value":["Master of Arts"]},{"label":"Description","value":"<p>Mismatch repair (MMR) is a crucial process that corrects errors produced by DNA polymerase. During DNA replication, the polymerase can incorporate an incorrect base or slip on the primer-template junction, leading to base-pair mismatches and insertion-deletion loops (IDLs), respectively. In eukaryotes, MutS homologs Msh2-Msh6 and Msh2-Msh3 recognize these errors; Msh2-Msh6 binds base-pair mismatches and smaller IDLs, and Msh2-Msh3 binds larger IDLs. Following error recognition, downstream proteins are recruited to repair the errorcontaining DNA, thus promoting genomic stability. Notably, the actions of Msh2- Msh3 on DNA can also lead to genomic instability. During replication, the polymerase is likely to slip on triplet nucleotide repeats (TNRs) and form hairpin structures. Msh2-Msh3 is thought to stabilize these hairpins and promote TNR expansion, which has been associated with various neurological disorders such as Huntington’s disease. Surprisingly, Msh2-Msh6 does not appear to promote TNR expansion in most disease models and may protect against TNR contractions. Studying the mechanistic differences between Msh2-Msh3 and Msh2-Msh6 actions on TNR hairpin structures will help with understanding their differential effects on TNR instability. This project aims to understand how Msh2-Msh6 recognizes and responds to its cognate error-containing DNA substrates compared to TNR hairpins. Gel mobility shift assays confirmed that Msh2-Msh6 forms a tight complex with a CAG hairpin, similar to base-pair mismatches, leading to the hypothesis that Msh2- Msh6 recognizes the A:A mismatches within the TNR hairpin stem. To test this hypothesis, Msh2-Msh6 ATPase activity was compared with error-containing DNA and TNR hairpin structures. Pre-steady state ATPase measurements indicate that Msh2-Msh6 responds to a CAG hairpin structure like a mismatch. Thus, although Msh2-Msh6 binds CAG hairpins with high affinity, it may slide off in an abortive attempt to initiate repair and therefore does not stabilize this DNA structure like Msh2-Msh3. Further kinetic analysis of Msh2-Msh6 interactions with TNR hairpins, including the effects of nucleotides, and its subsequent interaction with Mlh1-Pms1 is necessary to test this prediction. Together with analogous studies of Msh2-Msh3 in the future, these results may help resolve why Msh2-Msh6 does not promote TNR expansion and suggest ways to prevent the development of TNR-based neurological disorders.</p>"},{"label":"Digital Identifier","value":"m2017-k97"},{"label":"Thesis Advisor(s)","value":["Hingorani, Manju"]},{"label":"Digital Object Identifier (DOI)","value":"https://doi.org/10.14418/wes01.2.159"},{"label":"Discipline","value":["Molecular Biology and Biochemistry"]},{"label":"Date","value":"May 2017"},{"label":"EDTF Date","value":"D:00 M:00 Y:2017"},{"label":"Extent","value":"vii, 122 leaves : illustrations (chiefly color) ; 29 cm"},{"label":"File Format","value":["application/pdf"]},{"label":"Genre","value":["masters theses"]},{"label":"Digital Origin","value":["born digital"]},{"label":"Type","value":["Text"]}],"description":"Investigating the intersection between DNA mismatch repair and Triplet nucleotide repeat 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