Identification of a physiological phosphorylation site of the herpes simplex virus 1-encoded protein kinase Us3 which regulates its optimal catalytic activity in vitro and influences its function in infected cells.

Journal of Virology
Akihisa KatoYasushi Kawaguchi

Abstract

Us3 is a serine/threonine protein kinase encoded by herpes simplex virus 1 (HSV-1). Here, we report the identification of a physiological Us3 phosphorylation site on serine at position 147 (Ser-147) which regulates its protein kinase activity in vitro. Moreover, mutation of this site influences Us3 function, including correct localization of the enzyme and induction of the usual morphological changes in HSV-1-infected cells. These conclusions are based on the following observations: (i) in in vitro kinase assays, a domain of Us3 containing Ser-147 was specifically phosphorylated by Us3 and protein kinase A, while a mutant domain in which Ser-147 was replaced with alanine was not; (ii) in vitro, alanine replacement of Ser-147 (S147A) in Us3 resulted in significant impairment of the kinase activity of the purified molecule expressed in a baculovirus system; (iii) phosphorylation of Ser-147 in Us3 tagged with the monomeric fluorescent protein (FP) VenusA206K (VenusA206K-Us3) from Vero cells infected with a recombinant HSV-1 encoding VenusA206K-Us3 was specifically detected using an antibody that recognizes phosphorylated serine or threonine residues with arginine at the -3 and -2 positions; and (iv) the S147A mutation influenced s...Continue Reading

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Citations

Jul 28, 2010·Journal of Virology·Matthew S WaltersSaul Silverstein
Nov 19, 2010·Journal of Virology·Thary JacobHerman W Favoreel
Jun 21, 2013·Journal of Virology·Yuhei MaruzuruYasushi Kawaguchi
Apr 15, 2011·Uirusu·Yasushi Kawaguchi
Nov 28, 2015·Viruses·Ian B HogueLynn W Enquist
Dec 15, 2010·Protein & Cell·Haitao GuoHongyu Deng
Sep 4, 2010·Mitochondrion·Akane Ohta, Yukihiro Nishiyama
Jul 21, 2009·Virus Research·Barbara J KellyRussell J Diefenbach
Nov 10, 2013·Microbiology and Immunology·Naoto KoyanagiYasushi Kawaguchi
Dec 18, 2009·The Journal of General Virology·Shaunivan L LabiukSylvia van Drunen Littel-van den Hurk
Oct 28, 2014·Microbiology and Immunology·Chyntia JasirwanYasuko Mori
Jul 7, 2017·Journal of Virology·Ryosuke KobayashiYasushi Kawaguchi
Apr 1, 2016·Journal of Virology·Akihisa KatoYasushi Kawaguchi
Apr 18, 2014·Journal of Virology·Yuhei MaruzuruYasushi Kawaguchi
Jun 19, 2015·Journal of Virology·Zhuoming LiuYasushi Kawaguchi

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