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SR proteins regulate V6 exon splicing of CD44 pre-mRNA

  • Loh, Tiing Jen (School of life Sciences, Gwangju Institute of Science and Technology) ;
  • Moon, Heegyum (School of life Sciences, Gwangju Institute of Science and Technology) ;
  • Jang, Ha Na (School of life Sciences, Gwangju Institute of Science and Technology) ;
  • Liu, Yongchao (School of life Sciences, Gwangju Institute of Science and Technology) ;
  • Choi, Namjeong (School of life Sciences, Gwangju Institute of Science and Technology) ;
  • Shen, Shengfu (Willston Northampton School) ;
  • Williams, Darren Reece (School of life Sciences, Gwangju Institute of Science and Technology) ;
  • Jung, Da-Woon (School of life Sciences, Gwangju Institute of Science and Technology) ;
  • Zheng, Xuexiu (School of life Sciences, Gwangju Institute of Science and Technology) ;
  • Shen, Haihong (School of life Sciences, Gwangju Institute of Science and Technology)
  • Received : 2016.07.21
  • Accepted : 2016.08.16
  • Published : 2016.11.30

Abstract

CD44 pre-mRNA includes 20 exons, of which exons 1-5 ($C_1-C_5$) and exons 16-20 ($C_6-C_{10}$) are constant exons, whereas exons 6-15 ($V_1-V_{10}$) are variant exons. $V_6$-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 $V_6$ splicing using overexpression and lentivirus-mediated shRNA treatment. Using a CD44 $V_6$ minigene, we demonstrate that increased SRSF3 and SRSF4 expression do not affect $V_6$ splicing, but increased expression of SRSF1, SRSF6 and SRSF9 significantly inhibit $V_6$ 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 $V_6$ specific primer, we identified that reduced SRSF2 expression significantly reduced the $V_6$ isoform, but increased $V_{6-10}$ and $V_{6,8-10}$ isoforms. Our results indicate that SR proteins are important regulatory proteins for CD44 $V_6$ splicing.

Keywords

CD44;Pre-mRNA splicing;SR protein;$V_6$ exon

Acknowledgement

Supported by : National Research Foundation (NRF) of Korea, Gwangju Institute of Science and Technology (GIST)

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