Presentation Title

Identification of RNA binding motifs of Ubp3 and Bre5

Faculty Mentor

Sarah Mitchell

Start Date

18-11-2017 2:15 PM

End Date

18-11-2017 3:15 PM

Location

BSC-Ursa Minor 58

Session

Poster 3

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

The bulk of regulation of gene expression occurs at the level of the messenger RNA (mRNA). This regulation is largely carried out by proteins that influence mRNA modification, localization, degradation, and translation. Misregulation of mRNA-protein interactions can lead to a variety of diseases including cancer and neurodegenerative diseases. Thus, studying the activities of mRNA binding proteins is an important area of research. Recently, 66 previously unknown mRNA binding proteins were identified in S. cerevisiae, including the Ubp3/Bre5 ubiquitin protease complex. This complex removes ubiquitin from proteins, protecting these proteins from degradation. It is unclear why a complex with this activity would bind to mRNA. These proteins may represent a novel connection between the pathways of gene expression and protein degradation.

To study the function of Ubp3/Bre5 in mRNA biology, it is important to identify the mRNAs bound by this complex. SELEX (Systematic Evolution of Ligands by EXponential enrichment) is an in vitro technique that has been used to identify the sequence preference of many RNA binding proteins. Using SELEX, we are working to identify RNA sequences that are likely to be regulated by Ubp3/Bre5. By comparing these sequences to sequences of S. cerevisiae mRNA, we will be able to identify mRNAs that are likely regulated by this complex. Additionally, RNA motifs identified will be critical in establishing assays to study the mechanisms and functional consequence of Ubp3/Bre5 interaction with mRNA. Overall this work will identify interactions between a novel mRNA regulator and its targets, opening a pathway for the identification of exciting new mechanisms of gene expression regulation.

Summary of research results to be presented

Ubp3 and Bre5 have been expressed in E. coli cells and purified using affinity chromatography. We are currently optimizing the SELEX procedure for use with these proteins.

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Nov 18th, 2:15 PM Nov 18th, 3:15 PM

Identification of RNA binding motifs of Ubp3 and Bre5

BSC-Ursa Minor 58

The bulk of regulation of gene expression occurs at the level of the messenger RNA (mRNA). This regulation is largely carried out by proteins that influence mRNA modification, localization, degradation, and translation. Misregulation of mRNA-protein interactions can lead to a variety of diseases including cancer and neurodegenerative diseases. Thus, studying the activities of mRNA binding proteins is an important area of research. Recently, 66 previously unknown mRNA binding proteins were identified in S. cerevisiae, including the Ubp3/Bre5 ubiquitin protease complex. This complex removes ubiquitin from proteins, protecting these proteins from degradation. It is unclear why a complex with this activity would bind to mRNA. These proteins may represent a novel connection between the pathways of gene expression and protein degradation.

To study the function of Ubp3/Bre5 in mRNA biology, it is important to identify the mRNAs bound by this complex. SELEX (Systematic Evolution of Ligands by EXponential enrichment) is an in vitro technique that has been used to identify the sequence preference of many RNA binding proteins. Using SELEX, we are working to identify RNA sequences that are likely to be regulated by Ubp3/Bre5. By comparing these sequences to sequences of S. cerevisiae mRNA, we will be able to identify mRNAs that are likely regulated by this complex. Additionally, RNA motifs identified will be critical in establishing assays to study the mechanisms and functional consequence of Ubp3/Bre5 interaction with mRNA. Overall this work will identify interactions between a novel mRNA regulator and its targets, opening a pathway for the identification of exciting new mechanisms of gene expression regulation.