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Nucleic Acids Research, 1990, Vol. 18, No. 1 143-149
© 1990

MOLECULAR BIOLOGY

Splicing of a C.elegans myosin pre-mRNA in a human nuclear extract

Stephen C. Ogg1,*, Philip Anderson2 and Marvin p. Wickens1

1Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin Madison, WI 53706, USA 2Department of Genetics, College of Agricultural and Life Sciences, University of Wisconsin Madison, WI 53706, USA

Received September 15, 1989. Accepted November 20, 1989.

Splicing of mammalian introns requires that the intron possess at least 80 nucleotides. This length requirement presumably reflects the constraints of accommodating multiple snRNPs simultaneously in the same intron. In the free-living nematode, C. elegans, introns typically are 45 to 55 nucleotides in length. In this report, we determine whether C. elegans introns can obviate the mammalian length requirement by virtue of their structure or sequence. We demonstrate that a 53 nucleotide intron from the unc-54 gene of C. elegans does not undergo splicing in a mammalian (HeLa) nuclear extract. However, insertion of 31 nucleotides of foreign, prokaryotic sequence into the same intron results in efficient splicing. The observed splicing proceeds by the same two-step mechanism observed with mammalian introns, and exploits the same 3' and 5' splice sites as are used in C. elegans. The branch point used lies in the inserted sequence. We conclude that C. elegans splicing components are either fewer in number or smaller than their mammalian counterparts.

* Present address: Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143, USA


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