Exploring RBM6 Overexpression: Impact on Cell Division and Gene Splicing in HeLa Cells

Abstract

The RNA-binding motif protein 6 (RBM6) serves as an alternative splicing factor and tumor suppressor gene initially identified as a gene mapped to a lung cancer tumor suppressor gene locus on chromosome 3p21. Sharing structural similarities with RBM5 and RBM10, such as their two RNA recognition motifs (RRM) domains involved in posttranscriptional gene expression, two zinc finger-like regions, a G-patch and an OCRE domain, as well as a YjbI homology domain not found in RBM5 or RBM10. RBM6 is a splicing factor or interacts with other splicing factors based on its localization to IGCs (interchromatin granule clusters), also known as nuclear speckles, nuclear domains enriched in pre-mRNA splicing factors located in the nucleoplasm of mammalian cells. Changes in RBM6 expression have been reported to alter alternative splicing, to alter the cloning efficiency of HeLa cells, and RBM6 was shown to bind preferentially to a consensus sequence. For example, one study showed that decreased expression of RBM6 caused increased exclusion of NUMB exon 9, the opposite of what is observed following depletion of RBM10. It is notable that these effects could only be observed when RBM6 was stably knocked down in cell lines. While changes in RBM6 expression impact pre-mRNA splicing and cell proliferation, its precise mechanism is unclear. We are creating RBM6 cell lines that overproduce the protein in HeLa cells, as well as studying the effect of transient expression of normal and mutated RBM6 proteins. We will determine if overexpression of RBM6 will promote, and whether this correlates with changes in the rate of cell division. Using mutations in the protein we will investigate how the overexpression of RBM6 promotes the exclusion of the gene NUMB exon 9 and alters the splicing of other pro-apoptotic genes.Cellular and Molecular Biolog