1. Academic Validation
  2. Key pituitary miRNAs mediate the expression of pig GHRHR splice variants by regulating splice factors

Key pituitary miRNAs mediate the expression of pig GHRHR splice variants by regulating splice factors

  • Int J Biol Macromol. 2022 May 31;208:208-218. doi: 10.1016/j.ijbiomac.2022.03.070.
Zhaohui Teng 1 Linlin Hao 1 Rui Yang 1 Jie Song 1 Zhaoguo Wang 1 Yingying Jiao 1 Jiayuan Fang 1 Shuo Zheng 1 Ze Ma 1 Xi Chen 1 Songcai Liu 1 Yunyun Cheng 2
Affiliations

Affiliations

  • 1 College of Animal Science, Jilin University, Changchun, Jilin 130062, China.
  • 2 NHC Key Laboratory of Radiobiology, College of Public Health, Jilin University, Changchun 130021, China. Electronic address: chengyy@jlu.edu.cn.
Abstract

The growth hormone releasing hormone receptor (GHRHR) is well documented in organism growth and its alternative splicing may generate multiple functional GHRHR splice variants (SVs). Our previous study has demonstrated the key pituitary miRNAs (let-7e and miR-328-5p) in pig regulated the expression of GHRHR SVs by directly targeting to them. And according to recent reports, the interplay between miRNA-based silencing of mRNAs and alternative splicing of pre-mRNAs is a crucial post-transcriptional mechanism. In this study, SF3B3 and CPSF4 were firstly excavated as the splice factors that involved in the formation of GHRHR SVs mediated by let-7e and miR-328-5p through the comparation of the expression relations of GHRHR SVs, let-7e/miR-328-5p and SF3B3/CPSF4 in pituitary tissues between Landrace pigs and BaMa pigs, as well as the prediction of the target relations of let-7e/miR-328-5p with SF3B3 and/or CPSF4. SF3B3 and CPSF4 targeted by let-7e and miR-328-5p were further verified by performing dual-luciferase reporter assays and detecting the expression of target transcripts. Then the RT-PCR, RT-qPCR and Western blot assays were used to confirm SF3B3 and CPSF4 were involved in the formation of the GHRHR SVs, and in this process, let-7e and miR-328-5p mediated GHRHR SVs by regulating SF3B3 and CPSF4. Finally, the target site of SF3B3 on pre-GHRHR was on the Exon 12 to Exon14, while CPSF4 acted on the other fragments of the pre-GHRHR, which were explored by dual-luciferase reporter system preliminarily. To the best of our knowledge, this paper is the first to report the miRNAs regulate GHRHR SVs indirectly by splice factors.

Keywords

GHRHR; Pituitary; Splice factors; Splice variants; miRNAs.

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