Role of N-Terminal Region in Modulating RNA Binding Activity of the La Module in Vertebrate LARP6

La-related protein 6 (LARP6) belongs to the evolutionarily conserved superfamily of La-related proteins (LARPs). The LARP6 member is less well known, with most previous work focusing on the RNA binding domain La Module. But previous studies have shown that the N-terminal region (NTR) of LARP6 appears to play a role in RNA binding activity and specificity, as well as being suspected of interacting with the La Module domain. To further explore the effect of the NTR on the overall RNA-binding function of the protein, we created five protein constructs with the zebrafish model containing the NTR and the La Module domain, by successive deletions in the NTR part of the amino acids. Each construct was tested in RNA binding experiments using electrophoretic mobility shift assays, against different RNA ligands polyA, polyU, polyC and endogenous stem-loop HsCOL1A1. Each construct discriminates between polyC and other ligands. Constructions 38-290 and isolated La Module exhibit a tighter binding to all homopolymer ligands. Each construct binds to HsCOL1A1 an order of magnitude more tightly. Our results support that NTR modulates RNA binding activity and ligand specificity. Interestingly, the binding becomes tighter for constructs 38-290 and La Module, which could indicate a specific role of these few amino acids in participating in RNA binding affinity and specificity.

Name thesis:degree_name

Master of Science

Level thesis:degree_level

Masters

Discipline thesis:degree_discipline

Biochemistry

Grantor

Texas State University

Year dc:date.issued

2023

Author dc:creator

• Canet, Mithilde

Advisor dc:contributor.advisor

• Lewis, Karen A.

Committee members dc:contributor.committeemember

• David, Wendi M.
• Kerwin, Sean Michael

• LARP6
• RNA binding
• NTR
• activity modulation
• EMSA
• La module

Language dc:language.iso

en

Handle dc:identifier.uri

https://hdl.handle.net/10877/19996

OAI identifier oai:identifier

oai:digital.library.txst.edu:10877/19996