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SR proteins regulate V6 exon splicing of CD44 pre-mRNA
tiing jen loh1, ha na jang1, yongchao liu1, namjeong choi1, shengfu shen2, darren reece williams1, da-woon jung1, xuexiu zheng1, haihong Shen1,*
1life sciences, gwangju institute of science and technology,
2willston northamptin school
Abstract
CD44 pre-mRNA includes 20 exons, among which exons1-5 (C1-C5) and exons16-20 (C6-C10) are constant exons, whereas exons 6-15 (V1-V10) are variant exons. V6 exon containing isoforms has been known to be implicated in tumor cell invasion and metastasis. In the present study, we performed SR protein screen for CD44 V6 splicing using overexpression and lentivirus-mediated shRNA treatment. Using CD44 V6 minigene, we demonstrate that increased SRSF3 and SRSF4 expression do not affect V6 splicing, but increased expression of SRSF1, SRSF6 and SRSF9 inhibit V6 splicing significantly. In addition, using constitutive exon specific primer set, we could not detect alteration of CD44 splicing after SR protein-targeting shRNA treatment. However, using V6 specific primer, we identified that reduced SRSF2 expression significantly reduced V6 isoform, but increased V6-10 and V6,7-10 isoforms. Our results indicate that SR proteins are important regulatory proteins for CD44 V6 splicing.
Abstract, Accepted Manuscript(in press) [Submitted on July 21, 2016, Accepted on August 16, 2016]
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