Analysis of exonic splicing enhancers in the mouse gonadotropin-releasing hormone (GnRH) gene

Jin Han, Jae Young Seong, Kyungjin Kim, Wolfgang Wuttke, Hubertus Jarry

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9 Citations (Scopus)

Abstract

We have previously reported that efficient removal of the first intron (intron A) of gonadotropin-releasing hormone (GnRH) pre-mRNA is the prerequisite event for post-transcriptional regulation. In the present study, using an in vitro HeLa splicing system, we examined the enhancing activities of exonic elements for GnRH pre-mRNA splicing. While not excised by exon 2 alone, intron A was efficiently excised when exon 3 and/or exon 4 was combined with exon 2, suggesting the presence of exonic splicing enhancers (ESEs) in exons 3 and 4. Purine-rich sequences located in the border of exons 2 and 3 (denoted ESE3) and in exon 4 (ESE4) revealed strong splicing enhancing activities. Mutation in ESE3 decreased pre-mRNA splicing, while mutation in purine-rich sequences in exon 2 did not. We further analyzed the functional activity of ESE4 by mutations or deletions of the ESE4 sequence that consists of three purine-repeats separated by two spacers and a putative hairpin constructing sequence. An UV cross-linking assay using the RNA sequence of ESE4 examined the presence of ESE4-specific binding proteins in the nuclear extracts from GT1 hypothalamic GnRH neurons. Collectively, this study indicates that a sequence context of ESE4 and its binding proteins may be crucially involved in enhanced GnRH pre-mRNA splicing. However, it should be further clarified as to which splicing factor(s) is responsible for ESE4-dependent GnRH pre-mRNA splicing.

Original languageEnglish
Pages (from-to)157-166
Number of pages10
JournalMolecular and Cellular Endocrinology
Volume173
Issue number1-2
DOIs
Publication statusPublished - 2001 Feb 22
Externally publishedYes

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Keywords

  • ESE
  • GnRH
  • Intron A
  • Pre-mRNA
  • Purine-rich sequence
  • Splicing
  • UV-crosslinking

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Endocrinology

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