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Title: Systematic study of sequence motifs for RNA trans splicing in Trypanosoma brucei
Authors: Siegel, T.N.
Tan, K.S.W. 
Cross, G.A.M.
Issue Date: Nov-2005
Source: Siegel, T.N., Tan, K.S.W., Cross, G.A.M. (2005-11). Systematic study of sequence motifs for RNA trans splicing in Trypanosoma brucei. Molecular and Cellular Biology 25 (21) : 9586-9594. ScholarBank@NUS Repository.
Abstract: mRNA maturation in Trypanosoma brucei depends upon trans splicing, and variations in trans-splicing efficiency could be an important step in controlling the levels of individual mRNAs. RNA splicing requires specific sequence elements, including conserved 5′ splice sites, branch points, pyrimidine-rich regions [poly(Y) tracts],3′ splice sites (3′SS), and sometimes enhancer elements. To analyze sequence requirements for efficient trans splicing in the poly(Y) tract and around the 3′SS, we constructed a luciferase-β-galactosidase double-reporter system. By testing ∼90 sequences, we demonstrated that the optimum poly(Y) tract length is ∼25 nucleotides. Interspersing a purely undine-containing poly(Y) tract with cytidine resulted in increased ironssplicing efficiency, whereas purines led to a large decrease. The position of the poly(Y) tract relative to the 3′SS is important, and an AC dinucleotide at positions -3 and -4 can lead to a 20-fold decrease in irons splicing. However, efficient irons splicing can be restored by inserting a second AG dinucleotide downstream, which does not function as a splice site but may aid in recruitment of the splicing machinery. These findings should assist in the development of improved algorithms for computationally identifying a 3′SS and help to discriminate noncoding open reading frames from true genes in current efforts to annotate the T. brucei genome. Copyright © 2005, American Society for Microbiology. All Rights Reserved.
Source Title: Molecular and Cellular Biology
ISSN: 02707306
DOI: 10.1128/MCB.25.21.9586-9594.2005
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