Abstract
Previous studies using the yeast two-hybrid assay (Y2H) have identified cyclin L1 (CCNL1) and Ewing sarcoma breakpoint region 1 protein (EWSR1) as being interacting partners of tuftelin-interacting protein 11 (TFIP11). All three proteins are functionally related to the spliceosome and involved in pre-mRNA splicing activities. The spliceosome is a dynamic ribonucleoprotein complex responsible for pre-mRNA splicing of intronic regions, and is composed of five small nuclear RNAs (snRNAs) and μ140 proteins. TFIP11 appears to play a role in spliceosome disassembly allowing for the release of the bound lariat-intron. The roles of CCNL1 and EWSR1 in the spliceosome are poorly understood. Using fluorescently-tagged proteins and confocal microscopy we show that TFIP11, CCNL1 and EWSR1 frequently co-localize to speckled nuclear domains. These data would suggest that all three proteins participate in a common cellular activity related to RNA splicing events.
Highlights
Pre-mRNA splicing is an essential step of gene expression in many eukaryotic genes
We reported the intracellular colocalization of endogenous tuftelin-interacting protein 11 (TFIP11) with fluorescently tagged cyclin L1 (CCNL1) and Ewing sarcoma breakpoint region 1 protein (EWSR1) fusion proteins
We previously reported the interaction between TFIP11 and CCNL1, as well as, TFIP11 and EWSR1 using Y2H assay [10]
Summary
Pre-mRNA splicing is an essential step of gene expression in many eukaryotic genes. The splicing activity is carried out by spliceosome, a multicomponent ribonucleoprotein complex containing five. A number of TFIP11-interacting proteins involved in RNA processing have been identified in a yeast two-hybrid (Y2H) screening of a mouse embryonic cDNA expression library [10]. These included cyclin L1 (CCNL1), DEAD box polypeptide 47. EWSR1 has been shown to interact with integral components of the transcriptional complex [17], the splicing factor 1 (SF1) [18] and the U1 snRNP-specific protein C (U1C) [19] These data imply that EWSR1 has dual activities that span gene transcription and RNA splicing [13]. Data presented here suggest that TFIP11, CCNL1 and EWSR1 each participate in a common cellular activity related to the events of RNA processing
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