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Molecular and functional analysis of the gene CG2943, the gene KIAA0090 ortholog in Drosophila melanogaster

Grant number: 11/16666-2
Support type:Scholarships in Brazil - Master
Effective date (Start): March 01, 2012
Effective date (End): March 31, 2013
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal Investigator:Enilza Maria Espreafico
Grantee:Carlos Antonio Couto Lima
Home Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

The KIAA0090 gene, located on chromosome 1 (1p36.13), encodes a highly conserved protein whose evidence suggests its involvement in stress response pathways. Preliminary data obtained in our laboratory have shown high expression of the KIAA0090 gene in strains of metastatic melanoma. Its ortholog in Drosophila melanogaster is noted under the CG2943. Our experiments showed that animals homozygous for the insertion of a P element (transposon) at the end 5'do gene showed lethality in late pupa stage. Morphological analysis during metamorphosis showed no obvious anatomical defects or developmental delays when compared to control animals. In another experiment subjects were mechanically removed from the pupal envelope at the time that should emerge, demonstrating that they were still alive, however, with serious defects in locomotion, making it impossible to emerge normally. Currently we have a strain of transgenic Drosophila containing a sequence specific RNA interference for CG2943. By using the modular expression of UAS-GAL4 expression becomes possible to run the double-stranded RNA in different cell types in predetermined stages. The directed expression of the transgene in muscle cells led to pupal lethality, which makes it interesting, since it reproduces the phenotype observed earlier. RT-PCR analysis in real time are currently being conducted in order to assess the degree of silencing in these animals. By using Drosophila hope to explore the molecular function of the gene CG2943, enabling the transfer of knowledge to the study area of human cancer.