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PMID: 12972637 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Compensatory evolution of a precursor messenger RNA secondary structure in the Drosophila melanogaster Adh gene.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 100 ·No. 20 ·2003-09-30 ·Pages 11499-504

Chen Y, Stephan W

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.

MeSH Terms
Alcohol Dehydrogenase/genetics Animals Base Sequence Drosophila melanogaster/enzymology,genetics Introns Mutagenesis, Site-Directed Nucleic Acid Conformation RNA Splicing RNA, Messenger/chemistry,genetics Reverse Transcriptase Polymerase Chain Reaction
Chemicals
RNA, Messenger Alcohol Dehydrogenase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Chen Ying
Department of Biology II, University of Munich, 80333 Munich, Germany.
Stephan Wolfgang
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.1932834100
Published
2003-09-30
Epub
2003-00-12
Pages
11499-504
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC208787
Subset
IM
Grants
NIGMS NIH HHS · GM-58404 · United States
External Links
PubMed source DOI: 10.1073/pnas.1932834100 PMC Full Text View journal details
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