ELAV inhibits 3'-end processing to promote neural splicing of ewg pre-mRNA.
The embryonic lethal abnormal visual system (ELAV) is a gene-specific regulator of alternative pre-mRNA processing in neurons of Drosophila. Here we define a functional in vivo binding site for ELAV in neurons through the development of a reporter gene system in transgenic animals in combination with in vitro binding assays. ELAV binds to erect wing (ewg) RNA 3' of a polyadenylation site in the terminal intron 6. At this polyadenylation site, ELAV inhibits 3'-end processing in vitro in a dose-dependent and sequence-specific manner, and ELAV binding is necessary in vivo to promote splicing of ewg intron 6. Further, the AAUAAA poly(A) complex recognition sequence, together with ELAV, is required to regulate neural 3' splice site choice in vivo. In addition, the use of segmentally labeled RNA substrates in UV cross-linking assays suggest that ELAV does not inhibit or redirect binding of cleavage factor dCstF64 at the regulated polyadenylation site on ewg RNA. These data indicate that binding of 3'-end processing factors, together with ELAV, can regulate alternative splicing.
Soller
Matthias
M
Department of Biology and Volen Center for Complex Systems, Brandeis University, Waltham, Massachusetts 02454, USA. msoller@brandeis.edu
White
Kalpana
K
D000241:Adenosine / Q000378:metabolism; D029721:Drosophila Proteins; D051959:ELAV Proteins; D017930:Genes, Reporter; D009474:Neurons / Q000378:metabolism; D009479:Neuropeptides / Q000235:genetics* / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D011108:Polymers / Q000378:metabolism; D039104:RNA 3' Polyadenylation Signals; D012322:RNA Precursors / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D012261:Ribonucleoproteins / Q000378:metabolism*; D014157:Transcription Factors
D029721:Drosophila Proteins; D051959:ELAV Proteins; C081334:EWG protein, Drosophila; D009479:Neuropeptides; D009687:Nuclear Proteins; D011108:Polymers; D012322:RNA Precursors; D012261:Ribonucleoproteins; D014157:Transcription Factors; C069424:polyadenosine; D000241:Adenosine