Diversification of Drosophila chloride channel gene by multiple posttranscriptional mRNA modifications.
We have identified and analyzed a Drosophila melanogaster gene that encodes a chloride channel subunit (DrosGluCl-alpha) previously shown to function as a glutamate-gated chloride channel in an in vitro expression system. Sequence analysis of several cDNAs corresponding to the gene revealed sequence diversity in their open reading frames at seven specific sites. Site-specific A-to-G variations between cDNA and genomic sequences, consistent with RNA editing, were detected at five nucleotide positions. In addition, sequence variations among cDNA clones consistent with alternative splicing of mRNA were found at two different sites. In the 5' region, two small adjacent exons, containing similar but distinct modular sequences, are alternatively incorporated into the mature mRNA. In the 3' region, alternative splicing generates a variant encoding a protein with four additional amino acids just upstream of the fourth transmembrane domain. Combinations of RNA editing and alternative splicing can lead to extensive diversification of transcripts. These results give the first example of RNA editing in neurotransmitter-gated chloride channel genes or of alternative splicing in a glutamate-gated chloride channel gene of Drosophila.
Semenov
E P
EP
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA.
Pak
W L
WL
D017398:Alternative Splicing / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018118:Chloride Channels / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D018076:DNA, Complementary / Q000302:isolation & purification; D004330:Drosophila / Q000235:genetics*; D005091:Exons / Q000235:genetics; D007438:Introns / Q000235:genetics; D015640:Ion Channel Gating / Q000502:physiology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000502:physiology; D018377:Neurotransmitter Agents / Q000378:metabolism; D017393:RNA Editing / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics*
D018118:Chloride Channels; D017931:DNA Primers; D018076:DNA, Complementary; D018377:Neurotransmitter Agents; D012333:RNA, Messenger