This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Scamborova, P. Articles by Steitz, J. A. Search for Related Content PubMed PubMed Citation Articles by Scamborova, P. Articles by Steitz, J. A.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, March 2004, p. 1855-1869, Vol. 24, No. 5
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.5.1855-1869.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

An Intronic Enhancer Regulates Splicing of the Twintron of Drosophila melanogaster prospero Pre-mRNA by Two Different Spliceosomes

Department of Molecular Biophysics and Biochemistry, Yale University Howard Hughes Medical Institute, New Haven, Connecticut 06536-9812

Received 12 September 2003/ Returned for modification 28 October 2003/ Accepted 18 November 2003

We have examined the alternative splicing of the Drosophila melanogaster prospero twintron, which contains splice sites for both the U2- and U12-type spliceosome and generates two forms of mRNA, pros-L (U2-type product) and pros-S (U12-type product). We find that twintron splicing is developmentally regulated: pros-L is abundant in early embryogenesis while pros-S displays the opposite pattern. We have established a Kc cell in vitro splicing system that accurately splices a minimal pros substrate containing the twintron and have examined the sequence requirements for pros twintron splicing. Systematic deletion and mutation analysis of intron sequences established that twintron splicing requires a 46-nucleotide purine-rich element located 32 nucleotides downstream of the U2-type 5' splice site. While this element regulates both splicing pathways, its alteration showed the severest effects on the U2-type splicing pathway. Addition of an RNA competitor containing the wild-type purine-rich element to the Kc extract abolished U2-type splicing and slightly repressed U12-type splicing, suggesting that a trans-acting factor(s) binds the enhancer element to stimulate twintron splicing. Thus, we have identified an intron region critical for prospero twintron splicing as a first step towards elucidating the molecular mechanism of splicing regulation involving competition between the two kinds of spliceosomes.

This article has been cited by other articles:

• Mount, S. M., Gotea, V., Lin, C.-F., Hernandez, K., Makalowski, W. (2007). Spliceosomal small nuclear RNA genes in 11 insect genomes. RNA 13: 5-14 [Abstract] [Full Text]  
• McNally, L. M., Yee, L., McNally, M. T. (2006). Heterogeneous Nuclear Ribonucleoprotein H Is Required for Optimal U11 Small Nuclear Ribonucleoprotein Binding to a Retroviral RNA-processing Control Element: IMPLICATIONS FOR U12-DEPENDENT RNA SPLICING. J. Biol. Chem. 281: 2478-2488 [Abstract] [Full Text]  
• Touchon, M., Arneodo, A., d'Aubenton-Carafa, Y., Thermes, C. (2004). Transcription-coupled and splicing-coupled strand asymmetries in eukaryotic genomes. Nucleic Acids Res 32: 4969-4978 [Abstract] [Full Text]