PMID:12972637

Citation	Chen, Y and Stephan, W (2003) Compensatory evolution of a precursor messenger RNA secondary structure in the Drosophila melanogaster Adh gene. Proc. Natl. Acad. Sci. U.S.A. 100:11499-504
Abstract	Evidence for the evolutionary maintenance of a hairpin structure possibly involved in intron processing had been found in intron 1 of the alcohol dehydrogenase gene (Adh) in diverse Drosophila species. In this study, the putative hairpin structure was evaluated systematically in Drosophila melanogaster by elimination of either side of the stem using site-directed mutagenesis. The effects of these mutations and the compensatory double mutant on intron splicing efficiency and ADH protein production were assayed in Drosophila melanogaster Schneider L2 cells and germ-line transformed adult flies. Mutations that disrupt the putative hairpin structure right upstream of the intron branch point were found to cause a significant reduction in both splicing efficiency and ADH protein production. In contrast, the compensatory double mutant that restores the putative hairpin structure was indistinguishable from the WT in both splicing efficiency and ADH level. It was also observed by mutational analysis that a more stable secondary structure (with a longer stem) in this intron decreases both splicing efficiency and ADH protein production. Implications for RNA secondary structure and intron evolution are discussed.
Links	PubMed PMC208787 Online version:10.1073/pnas.1932834100
Keywords	Alcohol Dehydrogenase/genetics; Animals; Base Sequence; Drosophila melanogaster/enzymology; Drosophila melanogaster/genetics; Introns; Mutagenesis, Site-Directed; Nucleic Acid Conformation; RNA Splicing; RNA, Messenger/chemistry; RNA, Messenger/genetics; Reverse Transcriptase Polymerase Chain Reaction
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Gene product	Qualifier	GO ID	GO term name	Evidence Code	with/from	Aspect	Notes	Status
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