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Nucleic Acids Research, 2001, Vol. 29, No. 17 3477-3486
© 2001 Oxford University Press

Kinetics and regulation of site-specific endonucleolytic cleavage of human IGF-II mRNAs

Erwin L. van Dijk, John S. Sussenbach and P. Elly Holthuizen*

University Medical Center Utrecht, Department of Physiological Chemistry, Stratenum, Universiteitsweg 100, 3584 CG Utrecht, The Netherlands

Human insulin-like growth factor II (IGF-II) mRNA can be cleaved at a specific site in its 4 kb long 3'-UTR. This yields a stable 3' cleavage product of 1.8 kb consisting of a 3'-UTR and a poly(A) tail and an unstable 5' cleavage product containing the IGF-II coding region. After cleavage, the 5' cleavage product is targeted to rapid degradation and consequently is no longer involved in IGF-II protein synthesis. Cleavage is therefore thought to provide an additional way to control IGF-II gene expression. In this paper the kinetics and the efficiency of cleavage of IGF-II mRNAs are examined. The cleavage efficiency of IGF-II mRNAs carrying four different leaders (L1–L4) is enhanced in the highly structured leaders L1 and L3. Additionally, under standard cell culture conditions cleavage is a slow process that only plays a limited role in destabilisation and translation of the IGF-II mRNAs. However, in human Hep3B cells and CaCo2 cells which express IGF-II endogenously, cleavage is upregulated 3–5-fold at high cell densities. Regulated endonucleolytic cleavage of IGF-II mRNAs is restricted to cells in which IGF-II expression is related to specific cell processes.

* To whom correspondence should be addressed. Tel: +31 30 2538966; Fax: +31 30 2539035; Email: p.holthuizen{at}med.uu.nl


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