title: Btf and TRAP150 have distinct roles in regulating subcellular mRNA distribution.
abstract: Transcription of protein-coding genes in mammalian cells is coordinated with pre-mRNA processing as well as the assembly and nuclear export of mRNPs. Btf (BCLAF1) and TRAP150 (THRAP3) were previously reported to associate with in vitro spliced mRNPs and also as a part of the spliceosome, suggesting they are involved in pre-mRNA processing. Btf and TRAP150 are serine-arginine-rich (SR) proteins with significant sequence similarity, but the extent of their functional overlap is not yet clear. We show that both Btf and TRAP150 localize at a constitutively active β-tropomyosin (BTM) reporter minigene locus in mammalian cells. Both proteins also localize at a U2OS 2-6-3 reporter gene locus in a RNA polymerase II (RNAPII) transcription-dependent manner. While Btf and TRAP150 showed some overlap with reporter RNA and other pre-mRNA processing factors at transcription loci, they showed the most precise overlap with the exon junction complex (EJC) protein Magoh. Since EJC components have roles in nuclear export, we examined nuclear/cytoplasmic mRNA distribution after Btf or TRAP150 knockdown. Btf depletion caused an increase of β-tropomyosin minigene reporter transcripts in the cytoplasm as well as global increase of endogenous polyadenylated RNA in the cytoplasm, while TRAP150 depletion did not. We provide evidence that Btf has functions distinct from TRAP150 in regulating the subcellular distribution of mRNAs in human cells.
authors.affiliation: Wright State University, Dayton, OH, USA.
authors.forename: Sapna
authors.initials: S
authors.lastname: Varia
authors.forename: Divya
authors.initials: D
authors.lastname: Potabathula
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Deng
authors.forename: Athanasios
authors.initials: A
authors.lastname: Bubulya
authors.forename: Paula A
authors.initials: PA
authors.lastname: Bubulya
mesh_terms: D001692:Biological Transport; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D017381:Spliceosomes / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014335:Tropomyosin / Q000235:genetics / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: C119386:BCLAF1 protein, human; D004268:DNA-Binding Proteins; C491494:MAGOH protein, human; D009687:Nuclear Proteins; D012322:RNA Precursors; D012333:RNA, Messenger; D012097:Repressor Proteins; D012261:Ribonucleoproteins; C475774:THRAP3 protein, human; D014157:Transcription Factors; D014335:Tropomyosin; D025521:Tumor Suppressor Proteins; D012319:RNA Polymerase II
keywords: Btf/BCLAF1; TRAP150/THRAP3; exon junction complex; pre-mRNA processing; transcription sites