Deletions at intervening sequence splice sites in the alcohol dehydrogenase gene ofDrosophila
Open Access
- 25 November 1982
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 10 (22) , 7261-7272
- https://doi.org/10.1093/nar/10.22.7261
Abstract
Two formaldehyde-induce, homozygous viable ADH-negative mutants, Adhfn4and Adhfn6, possess no material that cross-reacts with antibody directed gainst ADH, no mature mRNA of wild-type size, and greatly reduced amounts of RNA that hybridizes with an Adh probe. We have cloned the genomic DNA sequences from these mutants in bacteriophage λ Charon 4 and subcloned the Adh region into plasmid vector pBR327. Restriction analyses revealed one small deletion in each of these mutants and DNA sequencing showed that the splice junctions of he 65-base pair (bp) intervening sequence (IVS) were altered. Both cloned mutant Adh genes, as well as the wild-type gene, are capable of promoting correct specific transcription, initiation in HeLa cell nuclear extracts in vitro. We conclude that Adhfn4and Adhfn6 are defective in RNA processing. Our results provide evidence for the importance of the splice junction sequences in normal ADH RNA processing and stabilization in Drosophila. We also speculate that splicing of ADH RNA proceeds in a nonrandom manner: mutations in one of the intervening sequences appear to cause accumulation of a large ADH RNA containing at least one other IVSKeywords
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