Survey for FMR1 exon 12 splicing activating candidate proteins

• Grant number: 11/03964-5
• Support type: Scholarships in Brazil - Master
• Effective date (Start): August 01, 2011
• Effective date (End): January 31, 2013
• Field of knowledge: Biological Sciences - Biochemistry
• Principal Investigator: Luciana Amaral Haddad
• Grantee: Marcelo Valpeteris de Campos
• Home Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Alternative splicing is considered a key mechanism in the generation of transcriptomic and proteomic diversity in eukaryotes. With the ability to originate mRNA variants from a single gene, it is implicated in up to 80% of human brain expressed trancripts. Splicing is regulated both by accessory nucleotide sequences (cis elements) and various protein components (trans elements) that interact with each other, resulting in an alteration of affinity between the spliceosome and specific splice sites. The mature product of splicing is, therefore, a result of the combination of these interactions. A gene with fundamental roles in central nervous system development that is subject to alternative splicing is the Fragile X Mental Retardation (FMR1) gene. The research group in which this project is inserted has recently demonstrated that the inclusion of exon 12 in mature Fmr1 transcripts is regulated during cerebral cortex development in rats. Also, it has produced a mini-gene aimed at the study of Fmr1 exon 12 splicing regulation. In the present proposal, our aim is to search for Fmr1 exon 12 splicing activating proteins. For such, I intend to perform affinity precipitation assays between proteins from nuclear cerebral cortex extract and biotinylated transcripts, which will be synthesized in vitro from the mini-gene clone DNA and from a mutant possessing an intronic deletion of a segment containing splicing enhancer candidates. The precipitated proteins will be evaluated by mass spectrometry and comparative analyses between both conditions. (AU)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

CAMPOS, Marcelo Valpeteris de. Screening for candidate proteins to activate FMR1 exon 12 splicing. 2014. Master's Dissertation - Universidade de São Paulo (USP). Instituto de Biociências São Paulo.