Nucleic Acids Research, 2001, Vol. 29, No. 11 2292-2302
© 2001 Oxford University Press
Multiple features contribute to efficient constitutive splicing of an unusually large exon
Department of Microbiology and Immunology, 1Department of Pathology and Laboratory Medicine and 2The Lucille Parker Markey Cancer Center, University of Kentucky College of Medicine, Lexington, KY 40536, USA
Vertebrate internal exons are usually between 50 and 400 nt long; exons outside this size range may require additional exonic and/or intronic sequences to be spliced into the mature mRNA. The mouse polymeric immunoglobulin receptor gene has a 654 nt exon that is efficiently spliced into the mRNA. We have examined this exon to identify features that contribute to its efficient splicing despite its large size; a large constitutive exon has not been studied previously. We found that a strong 5' splice site is necessary for this exon to be spliced intact, but the splice sites alone were not sufficient to efficiently splice a large exon. At least two exonic sequences and one evolutionarily conserved intronic sequence also contribute to recognition of this exon. However, these elements have redundant activities as they could only be detected in conjunction with other mutations that reduced splicing efficiency. Several mutations activated cryptic 5' splice sites that created smaller exons. Thus, the balance between use of these potential sites and the authentic 5' splice site must be modulated by sequences that repress or enhance use of these sites, respectively. Also, sequences that enhance cryptic splice site use must be absent from this large exon.
* To whom correspondence should be addressed. Tel: +1 859 257 5478; Fax: +1 859 323 2094; Email: mlpete01{at}pop.uky.edu Present address:Shirley R. Bruce, Department of Immunology, MD Anderson Cancer Center, University of Texas at Houston, Houston, TX 77030, USA
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