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Nucleic Acids Research, 1992, Vol. 20, No. 19 5003-5009
© 1992

MOLECULAR BIOLOGY

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

Durk Meinsma, Wiep Scheper1, P.Elly Holthuizen1, J.Leo Van den Brande and John S. Sussenbach1,*

Wilhelmina Childrens' Hospital, State University of Utrecht PO Box 18009, 3501 CA Utrecht, The Netherlands 1Laboratory for Physiological Chemistry, State University of Utrecht Vondellaan 24a, 3521 GG Utrecht, The Netherlands

* To whom correspondence should be addressed

Received July 30, 1992. Revised August 24, 1992. Accepted August 24, 1992.

The human insulin-like growth factor II (IGF-II) gene constitutes a complex transcriptlonal 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 ß-globin gene. Apparently, the expressed hybrid ß-globln-IGF-II mRNA contains all the regulatory elements to confer site-specific endonucleolytic cleavage.


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