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Eukaryotic Cell, June 2007, p. 940-948, Vol. 6, No. 6
1535-9778/07/$08.00+0     doi:10.1128/EC.00059-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Functional Characterization of Spliceosomal Introns and Identification of U2, U4, and U5 snRNAs in the Deep-Branching Eukaryote Entamoeba histolytica ^,

Departments of Microbiology and Immunology and Internal Medicine, Stanford University School of Medicine, Stanford, California 94305-5124

Received 27 February 2007/ Accepted 17 April 2007

Pre-mRNA splicing is essential to ensure accurate expression of many genes in eukaryotic organisms. In Entamoeba histolytica, a deep-branching eukaryote, approximately 30% of the annotated genes are predicted to contain introns; however, the accuracy of these predictions has not been tested. In this study, we mined an expressed sequence tag (EST) library representing 7% of amoebic genes and found evidence supporting splicing of 60% of the testable intron predictions, the majority of which contain a GUUUGU 5' splice site and a UAG 3' splice site. Additionally, we identified several splice site misannotations, evidence for the existence of 30 novel introns in previously annotated genes, and identified novel genes through uncovering their spliced ESTs. Finally, we provided molecular evidence for the E. histolytica U2, U4, and U5 snRNAs. These data lay the foundation for further dissection of the role of RNA processing in E. histolytica gene expression.

Published ahead of print on 27 April 2007.

Supplemental material for this article may be found at http://ec.asm.org/.

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