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Identification of RNA sequences and structures involved in site-specific cleavage of IGF-II mRNAs

Published online by Cambridge University Press:  01 December 1998

ERWIN L. VAN DIJK ,
JOHN S. SUSSENBACH  and
P. ELLY HOLTHUIZEN
ERWIN L. VAN DIJK
Affiliation:
Laboratory for Physiological Chemistry, Utrecht University, Utrecht, The Netherlands
JOHN S. SUSSENBACH
Affiliation:
Laboratory for Physiological Chemistry, Utrecht University, Utrecht, The Netherlands
P. ELLY HOLTHUIZEN
Affiliation:
Laboratory for Physiological Chemistry, Utrecht University, Utrecht, The Netherlands
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Abstract

Insulin-like growth factor-II (IGF-II) mRNAs are subject to site-specific endonucleolytic cleavage in the 3′ untranslated region (UTR), rendering an unstable 5′ cleavage product containing the coding region and a very stable 3′ cleavage product of 1.8 kb consisting of the 3′-UTR sequence and the poly(A) tail. Previously, it was established that two widely separated elements in the 3′-UTR (elements I and II), that can form a duplex structure, are necessary and sufficient for cleavage. To further investigate the sequence and secondary structure requirements for cleavage, we have introduced a number of mutations around the cleavage site and assayed their effects on cleavage. Several recognition determinants involved in the endonucleolytic cleavage of IGF-II mRNAs were identified. Mutational analysis around the cleavage site revealed that cleavage is sequence specific and that the cleavage site must be in a single-stranded conformation to allow efficient cleavage. In addition, we have identified an accessory protein that specifically interacts with a stem-loop structure located 133 to 73 nt upstream of the cleavage site.

Keywords

3′-UTRendonucleolytic cleavageIGF-IIinsulin-like growth factor-IIRNA–protein interactionsRNA secondary structure
Type
Research Article
Information
RNA , Volume 4 , Issue 12 , December 1998 , pp. 1623 - 1635
Copyright
© 1998 RNA Society

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