PMID: 9547281Jun 20, 1998Paper

Structure-function analysis of the trypanosomatid spliced leader RNA

Nucleic Acids Research
Igor GoncharovShulamit Michaeli

Abstract

In trypanosomes, all mRNAs possess a spliced leader (SL) at their 5' end. SL is added to pre-mRNA via trans -splicing from a small RNA, the SL RNA. To examine structure-function aspects of the trypanosomatid SL RNA, an in vivo system was developed in the monogenetic trypanosomatid Leptomonas collosoma to analyze the function of chimeric and site-directed SL RNA mutants in trans -splicing. Stable cell lines expressing chimeric and mutated SL RNA from the authentic SL RNA regulatory unit were obtained. The chimeric RNA was expressed and assembled into an SL RNP particle, but could not serve as a substrate in splicing. Mutations in loop II and III of L.collosoma SL RNA formed the Y structure intermediate. In addition, a double SL RNA mutant in loop II, and positions 7 and 8 of the intron, also formed the Y structure intermediate, suggesting that these intron positions, although proposed to participate in the interaction of SL RNA with U5, may not be crucial for the first step of the trans -splicing reaction. A mutation in the exon located in loop I was not utilized in splicing, suggesting the importance of exon sequences for trans -splicing in trypanosomes. However, a double SL RNA mutant in loop II and exon position 31 was utiliz...Continue Reading

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Citations

Feb 13, 2001·Molecular and Biochemical Parasitology·P XuG A Buck
Aug 25, 2000·Molecular and Cellular Biology·D Evans, T Blumenthal
Jun 26, 1999·The Journal of Biological Chemistry·N R Sturm, D A Campbell

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