Novel deletion mutants that enhance a distant upstream 5' splice in the E3 transcription unit of adenovirus 2.
Region E3 of adenovirus is a "complex" transcription unit: i.e. different mRNAs and proteins arise by differential RNA 3' end selection and differential splicing of the primary transcript. We are using viable virus mutants to understand the controls that dictate the specificity and efficiency of the RNA processing signals. We describe a novel class of deletion mutations that enhance a natural 5' splice site located approximately 740 nucleotides (nt) upstream. In particular, deletions within nt 1691-2044 in the E3 transcription unit result in a 5-fold enhancement of the 5' splice site at nt 951 (as reflected in steady-state mRNA). The effect is specific, because the deletions do not affect the 5' splice site at nt 372, and because deletions within nt 2044-2214 do not affect either the 951 or the 372 5' splice sites. As a consequence of the enhanced splicing at the 951 5' site, synthesis of the major E3 mRNA and the major E3 protein (gp19K) are dramatically reduced. At least one of the natural 3' splice sites, located at nt 2157, is the recipient of the enhanced splicing at the 951 5' splice site. We conclude that sequences located within nt 1691-2044 affect (probably in cis) splicing at the 951 5' splice site. We speculate that nt 1691-2044 includes a splicing control region which functions to suppress splicing at the 951 5' splice site.
Deutscher
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Bhat
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Pursley
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Cladaras
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Wold
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D000260:Adenoviruses, Human / Q000235:genetics*; D001483:Base Sequence; D002872:Chromosome Deletion; D004262:DNA Restriction Enzymes; D005814:Genes, Viral; D006801:Humans; D007624:KB Cells / Q000378:metabolism; D009154:Mutation; D011061:Poly A / Q000235:genetics; D012313:RNA / Q000235:genetics; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic
D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D004262:DNA Restriction Enzymes