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Mol. Cell. Biol., 01 1995, 235-245, Vol 15, No. 1
W Scheper, D Meinsma, PE Holthuizen and JS Sussenbach
Human insulin-like growth factor II (IGF-II) mRNAs are subject to site-
specific endonucleolytic cleavage in the 3' untranslated region, leading to
an unstable 5' cleavage product containing the IGF-II coding region and a
very stable 3' cleavage product of 1.8 kb. This endonucleolytic cleavage is
most probably the first and rate-limiting step in degradation of IGF-II
mRNAs. Two sequence elements within the 3' untranslated region are required
for cleavage: element I, located approximately 2 kb upstream of the
cleavage site, and element II, encompassing the cleavage site itself. We
have identified a stable double-stranded RNA stem structure (delta G = -100
kcal/mol [418.4 kJ/mol]) that can be formed between element I and a region
downstream of the cleavage site in element II. This structure is conserved
among human, rat, and mouse mRNAs. Detailed analysis of the requirements
for cleavage shows that the relative position of the elements is not
essential for cleavage. Furthermore, the distance between the coding region
and the cleavage site does not affect the cleavage reaction. Mutational
analysis of the long-range RNA-RNA interaction shows that not only the
double-stranded character but also the sequence of the stable RNA stem is
important for cleavage.
Copyright © 1995, American Society for Microbiology
Long-range RNA interaction of two sequence elements required for endonucleolytic cleavage of human insulin-like growth factor II mRNAs
Laboratory for Physiological Chemistry, Utrecht University, The Netherlands.
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