Mutagenesis of the Sindbis virus nsP1 protein: effects on methyltransferase activity and viral infectivity

Virology
H L WangVictor Stollar

Abstract

It has been suggested that four amino acids which are absolutely conserved in th nsP1 nonstructural proteins encoded by togaviruses and in the homologous proteins encoded by plant viruses in the Sindbis virus (SV) superfamily may constitute a "methyltransferase motif." In the Sindbis virus nsP1 protein (540 amino acids) these four amino acids are represented by His39, Arg91, Asp94, and Tyr249. Earlier, in assays of methyltransferase (MTase) activity generated in SV-infected cells, we had shown that amino acid changes at positions 87 and 88 of SV nsP1 resulted in a 10-fold lower Km for S-adenosyl methionine, the methyl donor in MTase reactions. Using site-directed mutagenesis we now report the expression of nsP1 in Escherichia coli, and in the infectious clone of Sindbis virus, Toto/1101, in which His39, Arg91, Asp94, and Tyr249 were changed one at a time to Ala. We also expressed nsP1 with C-terminal deletions of varying size, as well as with internal deletions in the C-terminal portion of the protein, in E. coli. Changing His39, Arg91, Asp94, or Tyr249 to Ala led to a loss of both MTase activity and viral infectivity; however, changing Ile369 to Val, a conservative change in the carboxy-terminal half of nsP1, had no effect on ...Continue Reading

Citations

Apr 23, 1999·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·K W Buck
Jul 11, 2000·Journal of Virology·T AholaP Auvinen
Dec 3, 2003·Journal of Virology·Indra DéDorothea L Sawicki
Dec 15, 2005·Journal of Virology·Dorothea L SawickiStanley G Sawicki
Nov 16, 2011·Virology Journal·Zhu Wu-yang, Liang Guo-dong
Apr 28, 2010·PLoS Pathogens·Mickaël BouvetEtienne Decroly
May 20, 1999·Antiviral Research·A LampioL Kääriäinen
Jul 30, 2015·The Journal of General Virology·Jonathan C RuppRichard W Hardy
Jul 18, 2001·The Journal of General Virology·K N JohnsonL A Ball
Sep 5, 2002·Reviews in Medical Virology·Jonathan O RaynerKurt I Kamrud
Oct 14, 2016·BMC Evolutionary Biology·Andrew G KellyPeter A White
Aug 20, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Alicia HurtadoDaniel Meruelo
Apr 3, 2015·Journal of Virology·K J SokoloskiR W Hardy
Jan 17, 2014·Journal of Virology·Kristina A Stoermer BurrackThomas E Morrison
Nov 8, 1996·The Journal of Biological Chemistry·P LaakkonenL Kääriäinen
Apr 5, 2000·The Journal of Biological Chemistry·L VasiljevaL Kääriäinen
Feb 22, 2017·World Journal of Gastroenterology : WJG·Mohammad Khalid Parvez
Jan 20, 2018·Scientific Reports·Sameer KumarSoma Chattopadhyay
Mar 11, 2006·The Journal of General Virology·Sareen E GalbraithGregory J Atkins
Sep 16, 2020·Antimicrobial Agents and Chemotherapy·Kristina Kovacikova, Martijn J van Hemert

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