The role of RNF113A in fidelity and efficiency of splicing reactions

• Grant number: 16/17963-4
• Support Opportunities: Scholarships abroad - Research
• Start date: January 02, 2017
• End date: February 01, 2017
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Patricia Pereira Coltri
• Grantee: Patricia Pereira Coltri
• Host Investigator: Melissa Jurica
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: University of California, Santa Cruz (UC Santa Cruz), United States

Abstract

Splicing is an essential step for control of gene expression. The pre-RNAs are composed by intermediate sequences, introns, and coding sequences named exons. Splicing is the removal of introns and exon ligation in a mature mRNA sequence, and it is performed by the spliceosome. This is a ~2 MDa complex composed of 5 snRNAs (small nuclear RNAs) and more than 100 proteins. Assembly and activity of the spliceosome are dependent on several rearrangements between RNAs and proteins. RNF113A is one of the protein components involved in spliceosome maturation. We previously observed that RNF113A associates to U4, U5 and U6 snRNAs and to PRP19 and BRR2. Additionally, removal of RNF113A from nuclear extracts reduces the amount of U5 snRNAs associated to the complex. These results indicated RNF113A is important for rearrangements that ultimately lead to catalytic activation of the spliceosome. Interestingly, addition of recombinant RNF113A inhibited in vitro splicing reactions, probably as "dominant negative" effect. Despite its importance for spliceosome rearrangements, the activity of this protein in splicing reactions is poorly explored. This proposal aims to analyze the role of RNF113A in splicing efficiency by performed in vitro reactions using pre-RNAs with mutations in splicing sites and in the intron. These experiments will be important to understand the role of RNF113A in splicing reactions.