SR proteins regulate V6 exon splicing of CD44 pre-mRNA
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
CD44 pre-mRNA includes 20 exons, of which exons 1-5 (C1-C5) and exons 16-20 (C6-C10) are constant exons, whereas exons 6-15 (V1-V10) are variant exons. V6-exon-containing isoforms have been known to be implicated in tumor cell invasion and metastasis. In the present study, we performed a SR protein screen for CD44 V6 splicing using overexpression and lentivirus-mediated shRNA treatment. Using a CD44 V6 minigene, we demonstrate that increased SRSF3 and SRSF4 expression do not affect V6 splicing, but increased expression of SRSF1, SRSF6 and SRSF9 significantly inhibit V6 splicing. In addition, using a constitutive exon-specific primer set, we could not detect alterations of CD44 splicing after SR protein-targeting shRNA treatment. However, using a V6 specific primer, we identified that reduced SRSF2 expression significantly reduced the V6 isoform, but increased V6-10 and V6,8-10 isoforms. Our results indicate that SR proteins are important regulatory proteins for CD44 V6 splicing.
Abstract
CD44 pre-mRNA includes 20 exons, of which exons 1-5 (C1-C5) and exons 16-20 (C6-C10) are constant exons, whereas exons 6-15 (V1-V10) are variant exons. V6-exon-containing isoforms have been known to be implicated in tumor cell invasion and metastasis. In the present study, we performed a SR protein screen for CD44 V6 splicing using overexpression and lentivirus-mediated shRNA treatment. Using a CD44 V6 minigene, we demonstrate that increased SRSF3 and SRSF4 expression do not affect V6 splicing, but increased expression of SRSF1, SRSF6 and SRSF9 significantly inhibit V6 splicing. In addition, using a constitutive exon-specific primer set, we could not detect alterations of CD44 splicing after SR protein-targeting shRNA treatment. However, using a V6 specific primer, we identified that reduced SRSF2 expression significantly reduced the V6 isoform, but increased V6-10 and V6,8-10 isoforms. Our results indicate that SR proteins are important regulatory proteins for CD44 V6 splicing.
참고문헌 (29)
- Green MR, "Pre-mrna splicing", Annu Rev Genet, 20, pp.671~708 (1986)
- David CJ, "Alternative pre-mRNA splicing regulation in cancer: Pathways and programs unhinged", Genes Dev, 24, pp.2343~2364 (2010)
- Cooper TA, "Rna and disease", Cell, 136, pp.777~793 (2009)
- Kim E, "Insights into the connection between cancer and alternative splicing", Trends Genet, 24, pp.7~10 (2008)
- Busch A, "Evolution of sr protein and hnrnp splicing regulatory factors", Wiley Interdiscip Rev RNA, 3, pp.1~12 (2012)
- Han SP, "Functional diversity of the hnrnps: Past, present and perspectives", Biochem J, 430, pp.379~392 (2010)
- Herrlich P, "Cd44 acts both as a growth-and invasiveness-promoting molecule and as a tumor-suppressing cofactor", Ann N Y Acad Sci, 910, pp.106~118 (2000)
- Screaton GR, "The identification of a new alternative exon with highly restricted tissue expression in transcripts encoding the mouse pgp-1(cd44)homing receptor. Comparison of all 10 variable exons between mouse, human, and rat", J Biol Chem, 268, pp.12235~12238 (1993)
- Bennett KL, "Cd44isoforms containing exon v3 are responsible for the presentation of heparin-binding growth factor", J Cell Biol, 128, pp.687~698 (1995)
- Orian-Rousseau V, "Adhesion proteins meet receptors: A common theme?", Adv Cancer Res, 101, pp.63~92 (2008)
- Tremmel M, "A cd44v6peptide reveals a role of cd44 in vegfr-2 signaling and angiogenesis", Blood, 114, pp.5236~5244 (2009)
- Orian-Rousseau V, "Cd44 is required for two consecutive steps in hgf", c-met signaling, 2002, pp.16, 3074~3086
- Legg JW, "Identification and functional analysis of the ezrin-binding site in the hyaluronan receptor, cd44", Curr Biol, 8, pp.705~708 (1998)
- Boukis LA, "Ser", arg-rich protein-mediated communication between u1and u2 small nuclear ribonucleoprotein particles, 2004, pp.279, 29647~29653
- Shao W, "A u1-u2 snrnp interaction network during intron definition", Mol Cell Biol, 32, pp.470~478 (2012)
- Bai Y, "Control of 3'splice site choice in vivo by asf", sf2 and hnrnp a1, 1999, pp.27, 1126~1134
- Zhong XY, "Sr proteins in vertical integration of gene expression from transcription to RNA processing to translation", Mol Cell, 35, pp.1~10 (2009)
- Loh TJ, "Sc35 promotes splicing of the c5-v6-c6 isoform of cd44 pre-mRNA", Oncol Rep, 31, pp.273~279 (2014)
- Yan Mi, "DEPDC1 is a novel cell cycle related gene that regulates mitotic progression", BMB Reports, 48(7), pp.413~418 (2015), 생화학분자생물학회
- 한아림, "Transcriptional repression of ANGPT1 by histone H3K9 demethylase KDM3B", BMB Reports, 48(7), pp.401~406 (2015), 생화학분자생물학회
- Sun Q, "General splicing factor sf2", asf promotes alternative splicing by binding to an exonic splicing enhancer, 1993, pp.7, 2598~2608
- Chandler SD, "Rna splicing specificity determined by the coordinated action of RNA recognition motifs in sr proteins", Proc Natl Acad Sci U S A, 94, pp.3596~3601 (1997)
- Anko ML, "Global analysis reveals srp20-and srp75-specific mrnps in cycling and neural cells", Nat Struct Mol Biol, 17, pp.962~970 (2010)
- Pandit S, "Genome-wide analysis reveals sr protein cooperation and competition in regulated splicing", Mol Cell, 50, pp.223~235 (2013)
- Simard MJ, "Srp30c is a repressor of 3'splice site utilization", Mol Cell Biol, 22, pp.4001~4010 (2002)
- Han J, "Sr proteins induce alternative exon skipping through their activities on the flanking constitutive exons", Mol Cell Biol, 31, pp.793~802 (2011)
- Erkelenz S, "Position-dependent splicing activation and repression by sr and hnrnp proteins rely on common mechanisms", RNA, 19, pp.96~102 (2013)
- Bielli P, "Regulation of bcl-x splicing reveals a role for the polypyrimidine tract binding protein(ptbp1", hnrnp i)in alternative 5' splice site selection, 2014, pp.42, 12070~12081
- van Weering DH, "A PCR-based method for the analysis of human cd44 splice products", PCR Methods Appl, 3, pp.100~106 (1993)