Structure-function analysis of the mRNA cap methyltransferase of Saccharomyces cerevisiae.

The Journal of Biological Chemistry
S P Wang, S Shuman

Abstract

The Saccharomyces cerevisiae mRNA cap methylating enzyme is a 436-amino acid protein encoded by the essential ABD1 gene. To identify structural features of ABD1 required for enzyme function, we introduced alanine mutations at 19 positions within a 205-amino acid region of similarity to the methyltransferase domain of the vaccinia capping enzyme. Three new recessive lethal mutations, E170A, D194A, and R206A, were identified. Structure-function relationships were clarified by introducing conservative substitutions at Glu-170, Asp-194, and Arg-206, and at Tyr-254 (an essential residue identified previously). Alleles E170D and D194E were viable, whereas E170Q and D194N were lethal; hence, acidic side chains were critical at both positions. R206K was viable, suggesting that a basic residue sufficed. Y254S was lethal, whereas Y254F was viable, albeit slow growing; thus, an aromatic side chain was important. The ABD1 mutations that were deleterious in vivo elicited catalytic defects in vitro. By studying the effects of amino- and carboxyl-terminal deletions, we defined a fully active catalytic domain of ABD1 from residues 130 to 426. Residues 110-129 were dispensable for methyltransferase activity in vitro, but essential for function ...Continue Reading

References

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Citations

Jun 20, 1998·Nucleic Acids Research·B SchwerS Shuman
Oct 6, 2000·Genes & Development·S C SchroederD Bentley
Jun 29, 1999·Cold Spring Harbor Symposia on Quantitative Biology·S McCrackenD Bentley
May 24, 2003·Cold Spring Harbor Symposia on Quantitative Biology·S Shuman
Dec 5, 2008·Journal of Virology·Andrea M Murphy, Valery Z Grdzelishvili
Jun 14, 2003·Journal of Virology·Nayanendu SahaBeate Schwer
Dec 5, 1997·Proceedings of the National Academy of Sciences of the United States of America·S Shuman
Sep 2, 1997·Proceedings of the National Academy of Sciences of the United States of America·S P WangS Shuman
Nov 15, 2015·Expert Opinion on Therapeutic Patents·John Okombo, Kelly Chibale
Jan 18, 2006·FEBS Letters·Ines BenzaghouMartin Bisaillon
Jan 28, 2015·RNA Biology·Magdalena ByszewskaJanusz M Bujnicki
Dec 23, 2009·The Biochemical Journal·Victoria H Cowling
Feb 1, 2014·Journal of Genetics and Genomics = Yi Chuan Xue Bao·Jun Liu, Guifang Jia
Jan 21, 2004·Molecular Cell·Carme FabregaChristopher D Lima
Feb 18, 2004·Molecular Cell·Stephanie C SchroederDavid Bentley
Aug 3, 2002·Nature Reviews. Molecular Cell Biology·Stewart Shuman
Dec 13, 2005·Cell Biochemistry and Function·Li Liu
Aug 17, 2004·The Biochemical Journal·Isabelle BougieMartin Bisaillon
Jun 24, 2003·The Journal of Biological Chemistry·Martin Bisaillon, Isabelle Bougie
Nov 1, 2001·The Journal of Biological Chemistry·Stephane HausmannStewart Shuman
Dec 16, 1998·The Journal of Biological Chemistry·C K HoS Shuman
Mar 30, 2001·The Journal of Biological Chemistry·Y L ChiuT M Rana
Oct 18, 2000·The Journal of Biological Chemistry·B SchwerS Shuman
Aug 8, 1998·Journal of Virology·C H Gross, S Shuman
Oct 29, 1998·Biochemical and Biophysical Research Communications·T TsukamotoK Mizumoto

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