Isolation of a Site-specifically Modified RNA from an Unmodified Transcript

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2006 Feb 14

PMID 16473844

Citations 10

Authors

Ya-Ming Hou

Zhi Li

Howard Gamper

Affiliations

Soon will be listed here.

Abstract

Natural RNAs contain many base modifications that have specific biological functions. The ability to functionally dissect individual modifications is facilitated by the identification and cloning of enzymes responsible for these modifications, but is hindered by the difficulty of isolating site-specifically modified RNAs away from unmodified transcripts. Using the m1G37 and m1A58 methyl modifications of tRNA as two examples, we demonstrate that non-pairing base modifications protect RNAs against the DNA-directed RNase H cleavage. This provide a new approach to obtain homogeneous RNAs with site-specific base modifications that are suitable for biochemical and functional studies.

Citing Articles

Kinetic Analysis of tRNA Methyltransferases.

Hou Y, Masuda I Methods Enzymol. 2015; 560:91-116.

PMID: 26253967 PMC: 4860815. DOI: 10.1016/bs.mie.2015.04.012.

The temperature sensitivity of a mutation in the essential tRNA modification enzyme tRNA methyltransferase D (TrmD).

Masuda I, Sakaguchi R, Liu C, Gamper H, Hou Y J Biol Chem. 2013; 288(40):28987-96.

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Recognition of guanosine by dissimilar tRNA methyltransferases.

Sakaguchi R, Giessing A, Dai Q, Lahoud G, Liutkeviciute Z, Klimasauskas S RNA. 2012; 18(9):1687-701.

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Control of catalytic cycle by a pair of analogous tRNA modification enzymes.

Christian T, Lahoud G, Liu C, Hou Y J Mol Biol. 2010; 400(2):204-17.

PMID: 20452364 PMC: 2892103. DOI: 10.1016/j.jmb.2010.05.003.

The homotetrameric phosphoseryl-tRNA synthetase from Methanosarcina mazei exhibits half-of-the-sites activity.

Hauenstein S, Hou Y, Perona J J Biol Chem. 2008; 283(32):21997-2006.

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