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  Box H/ACA snoRNAs are preferred substrates for the trimethylguanosine synthase in the divergent unicellular eukaryote Trichomonas vaginalis.

Simoes-Barbosa, A., Chakrabarti, K., Pearson, M. D., Benarroch, D., Shuman, S., & Johnson, P. J. (2012). Box H/ACA snoRNAs are preferred substrates for the trimethylguanosine synthase in the divergent unicellular eukaryote Trichomonas vaginalis. RNA, 18(9), 1656-1665. doi:10.1261/rna.034249.112.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-000F-EFDE-4 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-822F-3
Genre: Journal Article

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Simoes-Barbosa, A., Author
Chakrabarti, K., Author
Pearson, M. D.1, Author              
Benarroch, D., Author
Shuman, S., Author
Johnson, P. J., Author
Affiliations:
1Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578576              

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Free keywords: Trichomonas vaginalis; snoRNA; capping; trimethylguanosine synthase; pseudouridine
 Abstract: The 2,2,7-trimethylguanosine caps of eukaryal snRNAs and snoRNA are formed by the enzyme Tgs1, which catalyzes sequential guanine-N2 methylations of m7G caps. Atypically, in the divergent unicellular eukaryote Trichomonas vaginalis, spliceosomal snRNAs lack a guanosine cap and the recombinant T. vaginalis trimethylguanosine synthase (TvTgs) produces only m2,7G in vitro. Here, we show by direct metabolic labeling that endogenous T. vaginalis RNAs contain m7G, m2,7G, and m2,2,7G caps. Immunodepletion of TvTgs from cell extracts and TvTgs add-back experiments demonstrate that TvTgs produces m2,7G and m2,2,7G caps. Expression of TvTgs in yeast tgs1D cells leads to the formation of m2,7G and m2,2,7G caps and complementation of the lethality of a tgs1D mud2D strain. Whereas TvTgs is present in the nucleus and cytosol of T. vaginalis cells, TMG-containing RNAs are localized primarily in the nucleolus. Molecular cloning of anti-TMG affinity-purified T. vaginalis RNAs identified 16 box H/ACA snoRNAs, which are implicated in guiding RNA pseudouridylation. The ensemble of new T. vaginalis H/ACA snoRNAs allowed us to predict and partially validate an extensive map of pseudouridines in T. vaginalis rRNA.

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Language(s): eng - English
 Dates: 2012-07-302012-09
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1261/rna.034249.112
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Title: RNA
Source Genre: Journal
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Pages: - Volume / Issue: 18 (9) Sequence Number: - Start / End Page: 1656 - 1665 Identifier: -