Novel trypanosomatid small nucleolar RNAs that guide methylation: their genome organization, expression and potential use to direct specific methylation on target RNA molecules

Trypanosomatids are the causative agent of several major parasitic diseases including African trypanosomiasis, American trypanosomiasis, and leishmaniasis. These parasites possess unique RNA-processing mechanisms including trans-splicing of pre-mRNA and RNA editing of mitochondrial transcripts. In t...

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Bibliographic Details
Published inThe Israel Medical Association journal Vol. 2 Suppl; pp. 58 - 62
Main Authors Xu, Y X, Liu, L, Michaeli, S
Format Journal Article
LanguageEnglish
Published Israel 01.07.2000
Subjects
Animals
Animals, Genetically Modified
Chromosome Mapping
Cloning, Molecular
Gene Expression Regulation
Genetic Engineering
Genome, Protozoan
Methylation
Mitochondria - metabolism
Ribose - metabolism
RNA
RNA Precursors - genetics
RNA Splicing - genetics
RNA, Protozoan - genetics
RNA, Ribosomal, 18S - genetics
RNA, Ribosomal, 5.8S - genetics
RNA, Small Nucleolar - genetics
Transcription, Genetic
Trypanosomatina - genetics
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Summary:Trypanosomatids are the causative agent of several major parasitic diseases including African trypanosomiasis, American trypanosomiasis, and leishmaniasis. These parasites possess unique RNA-processing mechanisms including trans-splicing of pre-mRNA and RNA editing of mitochondrial transcripts. In this study, we identified a trypanosomatid novel group of small nucleolar RNAs that belong to the box C/D snoRNA, which were shown to guide ribose methylation on rRNA. Three snoRNA genes were identified; snoRNA-2 carrying a single snoRNA and g2 and b2 coding for a single or multiple snoRNAs, respectively. Mapping of the methylation sites guided by snoRNA-2 using two different approaches suggest that snoRNA-2 has the potential to guide methylation on both 5.8S and 18S rRNAs. The trypanosomes follow the same guide-methylation rule established for yeast and for mammals. As a first attempt to change the methylation pattern of target RNAs, we generated transgenic parasites carrying the B2 and snoRNA-2, which were engineered to shift the methylation site on rRNA. Despite efficient expression of these tagged snoRNAs, the novel methylation site was not generated. However, efficient expression of tagged snoRNAs in transgenic parasites opens the possibility of engineering novel methylation sites on different target RNAs in vivo.
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ISSN:1565-1088