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Originally published In Press as doi:10.1074/jbc.M208070200 on August 27, 2002

J. Biol. Chem., Vol. 277, Issue 45, 42733-42740, November 8, 2002
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Upstream Elements Present in the 3'-Untranslated Region of Collagen Genes Influence the Processing Efficiency of Overlapping Polyadenylation Signals*

Barbara J. Natalizio, Luis C. Muñiz, George K. ArhinDagger , Jeffrey WiluszDagger , and Carol S. Lutz§

From the Department of Biochemistry and Molecular Biology and Dagger  Department of Microbiology and Molecular Genetics, University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark, New Jersey 07103

3'-Untranslated regions (UTRs) of genes often contain key regulatory elements involved in gene expression control. A high degree of evolutionary conservation in regions of the 3'-UTR suggests important, conserved elements. In particular, we are interested in those elements involved in regulation of 3' end formation. In addition to canonical sequence elements, auxiliary sequences likely play an important role in determining the polyadenylation efficiency of mammalian pre-mRNAs. We identified highly conserved sequence elements upstream of the AAUAAA in three human collagen genes, COL1A1, COL1A2, and COL2A1, and demonstrate that these upstream sequence elements (USEs) influence polyadenylation efficiency. Mutation of the USEs decreases polyadenylation efficiency both in vitro and in vivo, and inclusion of competitor oligoribonucleotides representing the USEs specifically inhibit polyadenylation. We have also shown that insertion of a USE into a weak polyadenylation signal can enhance 3' end formation. Close inspection of the COL1A2 3'-UTR reveals an unusual feature of two closely spaced, competing polyadenylation signals. Taken together, these data demonstrate that USEs are important auxiliary polyadenylation elements in mammalian genes.

* This work was funded by American Cancer Society Grant RPG-00-265-01-GMC, Arthritis Investigator Award 2AI-LUT-A-5, Arthritis Foundation, New Jersey Chapter Grant 3AI-LUT-A (to C. S. L.), and National Institutes of Health Grants CA80062 and GM63832 (to J. W.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ To whom correspondence should be addressed: MSB E671, 185 S. Orange Ave., UMDNJ-NJMS, Newark, NJ 07103. Tel.: 973-972-0899; Fax: 973-972-5594; E-mail: lutzcs@umdnj.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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