Construction and characterization of thermosensitive mutants of the general splicing-factor Cwc24 in Saccharomyces cerevisiae

• Grant number: 20/03315-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2020
• End date: December 31, 2020
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Carla Columbano de Oliveira
• Grantee: Moises Ricardo de Almeida Barros
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 15/06477-9 - Functional study of the components of the Saccharomyces cerevisiae exosome and the spliceosome, and their role in posttranscriptional control of gene expression, AP.TEM

Abstract

Splicing is a maturation process of pre-mRNAs, essential in eukaryotes, through which intron sequences are extracted from the precursor mRNAs, allowing the ligation of the exons and the formation of the protein coding sequences. Over a hundred factors are involved in this process, and among them, the protein Cwc24, which binds the intron 5'-end, protecting it from premature nucleophilic attacks. Cwc24 binds transiently to pre-mRNA and to the spliceosome complex and the mechanisms controlling its binding and later release has not been characterized. The aim of this project is the construction of aleatory mutants of Cwc24 that may affect its affinity for the spliceosome. Splicing is a maturation process of pre-mRNAs, essential in eukaryotes, through which intron sequences are extracted from the precursor mRNAs, allowing the ligation of the exons and the formation of the protein coding sequences. Over a hundred factors are involved in this process, and among them, the protein Cwc24, which binds the intron 5'-end, protecting it from premature nucleophilic attacks. Cwc24 binds transiently to pre-mRNA and to the spliceosome complex and the mechanisms controlling its binding and later release has not been characterized. The aim of this project is the construction of aleatory mutants of Cwc24 that may affect its affinity for the spliceosome.