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Nucleic Acids Res. 1992 Oct 11;20(19):5003-9. doi: 10.1093/nar/20.19.5003.

Site-specific cleavage of IGF-II mRNAs requires sequence elements from two distinct regions of the IGF-II gene.

Nucleic acids research

D Meinsma, W Scheper, P E Holthuizen, J L Van den Brande, J S Sussenbach

Affiliations

  1. Wilhelmina Childrens' Hospital, State University of Utrecht, The Netherlands.

PMID: 1408818 PMCID: PMC334276 DOI: 10.1093/nar/20.19.5003
Free PMC Article

Abstract

The human insulin-like growth factor II (IGF-II) gene constitutes a complex transcriptional unit that contains nine exons and four promoters. Expression of the IGF-II gene yields a family of mRNAs that all encode prepro-IGF-II. In addition, a stable 1.8 kb RNA is formed that is derived from the 3' untranslated region of exon 9. Recently, we have shown that this RNA species arises by site-specific endonucleolytic cleavage of IGF-II mRNAs and not by transcription from a separate promoter. In the present study we establish that two widely separated sequence elements of approximately 300 nucleotides, both located within exon 9, are required for this cleavage reaction. The first element encompasses about 200 nucleotides upstream and 100 nucleotides downstream of the cleavage site, while the second element is located within a region of 330 nucleotides about 2 kb upstream of the cleavage site. Interestingly, site-specific cleavage also occurred when a fragment from exon 9 of the IGF-II gene containing these two elements was inserted into the 3' untranslated part of the beta-globin gene. Apparently, the expressed hybrid beta-globin-IGF-II mRNA contains all the regulatory elements to confer site-specific endonucleolytic cleavage.

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