The open reading frame TTC1157 of Thermus thermophilus HB27 encodes the methyltransferase forming N2-methylguanosine at position 6 in tRNA
- 15 February 2012
- journal article
- Published by Cold Spring Harbor Laboratory in RNA
- Vol. 18 (4), 815-824
- https://doi.org/10.1261/rna.030411.111
Abstract
N2-methylguanosine (m2G) is found at position 6 in the acceptor stem of Thermus thermophilus tRNAPhe. In this article, we describe the cloning, expression, and characterization of the T. thermophilus HB27 methyltransferase (MTase) encoded by the TTC1157 open reading frame that catalyzes the formation of this modified nucleoside. S-adenosyl-L-methionine is used as donor of the methyl group. The enzyme behaves as a monomer in solution. It contains an N-terminal THUMP domain predicted to bind RNA and contains a C-terminal Rossmann-fold methyltransferase (RFM) domain predicted to be responsible for catalysis. We propose to rename the TTC1157 gene trmN and the corresponding protein TrmN, according to the bacterial nomenclature of tRNA methyltransferases. Inactivation of the trmN gene in the T. thermophilus HB27 chromosome led to a total absence of m2G in tRNA but did not affect cell growth or the formation of other modified nucleosides in tRNAPhe. Archaeal homologs of TrmN were identified and characterized. These proteins catalyze the same reaction as TrmN from T. thermophilus. Individual THUMP and RFM domains of PF1002 from Pyrococcus furiosus were produced. These separate domains were inactive and did not bind tRNA, reinforcing the idea that the THUMP domain acts in concert with the catalytic domain to target a particular position of the tRNA molecule.Keywords
This publication has 50 references indexed in Scilit:
- Crystal structures of the tRNA:m 2 G6 methyltransferase Trm14/TrmN from two domains of lifeNucleic Acids Research, 2012
- Formation of m 2 G6 in Methanocaldococcus jannaschii tRNA catalyzed by the novel methyltransferase Trm14Nucleic Acids Research, 2011
- Pseudouridine at position 55 in tRNA controls the contents of other modified nucleotides for low-temperature adaptation in the extreme-thermophilic eubacterium Thermus thermophilusNucleic Acids Research, 2010
- Trm112p Is a 15-kDa Zinc Finger Protein Essential for the Activity of Two tRNA and One Protein Methyltransferases in YeastPublished by Elsevier BV ,2010
- N 7-Methylguanine at position 46 (m7G46) in tRNA from Thermus thermophilus is required for cell viability at high temperatures through a tRNA modification networkNucleic Acids Research, 2009
- Aquifex aeolicus tRNA (N2,N2-Guanine)-dimethyltransferase (Trm1) Catalyzes Transfer of Methyl Groups Not Only to Guanine 26 but Also to Guanine 27 in tRNAPublished by Elsevier BV ,2009
- Functional specialization of domains tandemly duplicated within 16S rRNA methyltransferase RsmCNucleic Acids Research, 2007
- Ribosomal RNA guanine-(N2)-methyltransferases and their targetsNucleic Acids Research, 2007
- Methyltransferase That Modifies Guanine 966 of the 16 S rRNAPublished by Elsevier BV ,2007
- THUMP from archaeal tRNA:m22G10 methyltransferase, a genuine autonomously folding domainNucleic Acids Research, 2006