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Characterization of the role to the general splicing-factor CWC24 in pre-mRNA in Saccharomyces cerevisiae

Grant number: 21/14137-4
Support Opportunities:Scholarships in Brazil - Doctorate (Direct)
Effective date (Start): January 01, 2022
Status:Discontinued
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:20/00901-1 - Posttranscriptional control of gene expression: pre-rRNA processing, mRNA degradation, splicing and snoRNP assembly in Saccharomyces cerevisiae, AP.TEM
Associated scholarship(s):23/16763-5 - CHARACTERIZATION OF THE ROLE OF THE SPLICING-FACTOR CWC24 IN pre-RNA IN Saccharomyces cerevisiae, BE.EP.DD

Abstract

All RNAs transcribed in eukaryotic cells undergo maturation stages. One of the steps that pre-mRNAs go through is the splicing process, where the non-coding sequences (introns) are removed and the coding sequences (exons) joined. For this process to occur, two transesterification reactions are necessary, which in turn are mediated by a multimeric complex called a spliceosome, composed of five snRNPs (small nuclear ribonucleoproteins), U1, U2, U4, U5 and U6, and several proteins. The spliceosome assembly occurs during the splicing process, requiring several conformational changes of the subcomplexes, so that two transesterification reactions occur during the splicing. These conformational changes and the composition of the complex are mediated by the action of helicases and other ATPases, and consequently, involve ATP consumption. In addition to the snRNPs and ATPase complexes, the pre-mRNA splicing requires the NTC complex (NineTeen Complex), which participates in the two transesterification reactions, with its composition changing throughout the process. Among the proteins that make up the NTC complex, we highlight Cwc24, which transiently binds to the spliceosome and has the function of interacting with the 5' end of the intron to prevent the first premature transesterification reaction, which in turn would lead to a non-functional mRNA. Thus, this project proposes the characterization of the mechanisms that lead to the association and disassociation of Cwc24 from the spliceosome. (AU)

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