Using ubiquitin variants to understand the function of RFWD3 at the molecular level
Standing
Undergraduate
Type of Proposal
Oral Research Presentation
Challenges Theme
Open Challenge
Your Location
Windsor, Ontatrio
Faculty
Faculty of Science
Faculty Sponsor
Yufeng Tong
Proposal
Tagging of cellular proteins with marks, also known as post-translational modification, is a natural mechanism that controls the function of proteins and biological pathways. Among all protein modifications, ubiquitination is the second most abundant. It involves the attachment of ubiquitin, a small protein, unto a target protein. Once ubiquitinated, the structure of the target protein is drastically altered, resulting in a different fate. Ubiquitination is critical to DNA damage repair. Our research uses engineered ubiquitin mutants, known as ubiquitin variants (UbVs), to study the function of a ubiquitination enzyme, RFWD3, that is important for DNA damage repair. UbVs bind with the target protein they raised against tightly and specifically, thus perturb the activity of that enzyme and reveal its function. Heritable mutations that negatively affect the function of RFWD3 have been linked to a rare cancer known as Fanconi anemia (FA); making this a potential therapeutic target. Using UbVs specific for RFWD3 we aim to understand how it recognizes substrates and to elucidate the molecular details of its involvement in DNA damage repair. By screening a candidate library of UbVs engineered to bind the substrate-binding domain of RFWD3 we will identify the best binders for further analyses in cancer cells. Ultimately, the UbVs identified in this work have the potential to become a protein-based drug to treating DNA damage repair-related diseases.
Using ubiquitin variants to understand the function of RFWD3 at the molecular level
Tagging of cellular proteins with marks, also known as post-translational modification, is a natural mechanism that controls the function of proteins and biological pathways. Among all protein modifications, ubiquitination is the second most abundant. It involves the attachment of ubiquitin, a small protein, unto a target protein. Once ubiquitinated, the structure of the target protein is drastically altered, resulting in a different fate. Ubiquitination is critical to DNA damage repair. Our research uses engineered ubiquitin mutants, known as ubiquitin variants (UbVs), to study the function of a ubiquitination enzyme, RFWD3, that is important for DNA damage repair. UbVs bind with the target protein they raised against tightly and specifically, thus perturb the activity of that enzyme and reveal its function. Heritable mutations that negatively affect the function of RFWD3 have been linked to a rare cancer known as Fanconi anemia (FA); making this a potential therapeutic target. Using UbVs specific for RFWD3 we aim to understand how it recognizes substrates and to elucidate the molecular details of its involvement in DNA damage repair. By screening a candidate library of UbVs engineered to bind the substrate-binding domain of RFWD3 we will identify the best binders for further analyses in cancer cells. Ultimately, the UbVs identified in this work have the potential to become a protein-based drug to treating DNA damage repair-related diseases.