Document Type
Article
Publication Date
2007
Source Publication
Nucleic Acids Research
Source ISSN
0305-1048
Abstract
In Saccharomyces cerevisiae, a two-subunit methyltransferase (Mtase) encoded by the essential genes TRM6 and TRM61 is responsible for the formation of 1-methyladenosine, a modified nucleoside found at position 58 in tRNA that is critical for the stability of . The crystal structure of the homotetrameric m1A58 tRNA Mtase from Mycobacterium tuberculosis, TrmI, has been solved and was used as a template to build a model of the yeast m1A58 tRNA Mtase heterotetramer. We altered amino acids in TRM6 and TRM61 that were predicted to be important for the stability of the heteroligomer based on this model. Yeast strains expressing trm6 and trm61 mutants exhibited growth phenotypes indicative of reduced m1A formation. In addition, recombinant mutant enzymes had reduced in vitro Mtase activity. We demonstrate that the mutations introduced do not prevent heteroligomer formation and do not disrupt binding of the cofactor S-adenosyl-l-methionine. Instead, amino acid substitutions in either Trm6p or Trm61p destroy the ability of the yeast m1A58 tRNA Mtase to bind , indicating that each subunit contributes to tRNA binding and suggesting a structural alteration of the substrate-binding pocket occurs when these mutations are present.
Recommended Citation
Ozanick, Sarah G.; Bujnicki, Janusz M.; Sem, Daniel S.; and Anderson, James T., "Conserved Amino Acids in Each Subunit of the Heteroligomeric tRNA m1A58 Mtase from Saccharomyces cerevisiae Contribute to tRNA Binding" (2007). Chemistry Faculty Research and Publications. 394.
https://epublications.marquette.edu/chem_fac/394
Comments
Published version. Nucleic Acids Research, Vol. 35, No. 20 (2007): 6808-6819. DOI. © 2007 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License.