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Splicing in Caenorhabditis elegans does not require an AG at the 3' splice acceptor site

Aroian, Raffi V. and Levy, Adam D. and Koga, Makoto and Ohshima, Yasumi and Kramer, James M. and Sternberg, Paul W. (1993) Splicing in Caenorhabditis elegans does not require an AG at the 3' splice acceptor site. Molecular and Cellular Biology, 13 (1). pp. 626-637. ISSN 0270-7306. http://resolver.caltech.edu/CaltechAUTHORS:AROmcb93

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Abstract

The dinucleotide AG, found at the 3' end of virtually all eukaryotic pre-mRNA introns, is thought to be essential for splicing. Reduction-of- function mutations in two Caenorhabditis elegans genes, the receptor tyrosine kinase gene let-23 and the collagen gene dpy-10, both alter the AG at the end of a short (ca. 50-nucleotide) intron to AA. The in vivo effects of these mutations were studied by sequencing polymerase chain reaction-amplified reverse-transcribed RNA isolated from the two mutants. As expected, we find transcripts that splice to a cryptic AG, skip an exon, and retain an unspliced intron. However, we also find significant levels of splicing at the mutated 3' splice site (AA) and at nearby non-AG dinucleotides. Our results indicate that for short C. elegans introns an AG is not required for splicing at either the correct 3' splice site or incorrect sites. Analysis of a splice site mutant involving a longer, 316-nucleotide C. elegans intron indicates that an AG is also not required there for splicing. We hypothesize that elements besides the invariant AG, e.g., an A-U-rich region, a UUUC motif, and/or a potential branch point sequence, are directing the selection of the 3' splice site and that in wild-type genes these elements cooperate so that proper splicing occurs.


Item Type:Article
Additional Information:Copyright © 1993, American Society for Microbiology Received 5 May 1992/Returned for modification 5 August 1992/Accepted 28 October 1992 We thank Gregg Jongeward for the mutant let-23(sy97sy122), Tom Blumenthal for discussions on splicing in C. elegans, and Bob Horvitz for suggesting promiscuous splicing as a mechanism for generating diversity. We thank James Deshler, Andy Golden, Paul Kayne, and John Rossi for discussions and comments. We thank Jeff Miner for help with RNase protection and Sean Tavtigian and the Dervan Laboratory for help with Phosphorlmager quantitation. We thank Jane Mendel for wild-type RNA, Mike Krause for plasmid pT7/T3-18-103, Joseph LaRusso for help with sequencing, and Paul Kayne, Russell Hill, and Giovanni Lesa for technical advice. P.W.S. is an investigator of the Howard Hughes Medical Institute. This research was supported by grants HD23690 to P.W.S. and HD22028 to J.M.K.
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Deposited On:13 Feb 2006
Last Modified:26 Dec 2012 08:45

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