Site-Directed Mutagenesis of the REV1 BRCT Domain

Presenter's Name(s)

Hannah Koval

Conference Year

2024

Abstract

The translesion synthesis (TLS) polymerase REV1 modulates error-prone replication of DNA and contains a BRCT domain that interacts with proliferating cell nuclear antigen (PCNA). Structural analysis has identified five amino acid residues within the BRCT domain that are critical to the BRCT-PCNA interaction. Site-directed mutagenesis was used to generate point mutants for the five amino acid residues and Sanger sequencing confirmed successful generation of the R68A, G76R, Y81A, and K86A point mutants. These point mutants provide a useful tool for investigating the in vitro significance of the BRCT-PCNA interaction on TLS.

Primary Faculty Mentor Name

Nimrat Chatterjee

Status

Undergraduate

Student College

College of Arts and Sciences

Program/Major

Biochemistry

Primary Research Category

Life Sciences

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Site-Directed Mutagenesis of the REV1 BRCT Domain

The translesion synthesis (TLS) polymerase REV1 modulates error-prone replication of DNA and contains a BRCT domain that interacts with proliferating cell nuclear antigen (PCNA). Structural analysis has identified five amino acid residues within the BRCT domain that are critical to the BRCT-PCNA interaction. Site-directed mutagenesis was used to generate point mutants for the five amino acid residues and Sanger sequencing confirmed successful generation of the R68A, G76R, Y81A, and K86A point mutants. These point mutants provide a useful tool for investigating the in vitro significance of the BRCT-PCNA interaction on TLS.