Phosphorylated tyrosine 93 of hepatitis C virus nonstructural protein 5A is essential for interaction with host c-Src and efficient viral replication.
NS5A
Src
Src homology 2 domain (SH2 domain)
hepatitis C virus (HCV)
liver disease
nonstructural protein 5A
phosphotyrosine
viral replication
virion assembly
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
03 05 2019
03 05 2019
Historique:
received:
22
01
2019
revised:
11
03
2019
pubmed:
14
3
2019
medline:
8
1
2020
entrez:
14
3
2019
Statut:
ppublish
Résumé
The hepatitis C virus (HCV) nonstructural protein 5A (NS5A) plays a key role in viral replication and virion assembly, and the regulation of the assembly process critically depends on phosphorylation of both serine and threonine residues in NS5A. We previously identified SRC proto-oncogene, nonreceptor tyrosine kinase (c-Src), as an essential host component of the HCV replication complex consisting of NS5A, the RNA-dependent RNA polymerase NS5B, and c-Src. Pulldown assays revealed an interaction between NS5A and the Src homology 2 (SH2) domain of c-Src; however, the precise binding mode remains undefined. In this study, using a variety of biochemical and biophysical techniques, along with molecular dynamics simulations, we demonstrate that the interaction between NS5A and the c-Src SH2 domain strictly depends on an intact phosphotyrosine-binding competent SH2 domain and on tyrosine phosphorylation within NS5A. Detailed analysis of c-Src SH2 domain binding to a panel of phosphorylation-deficient NS5A variants revealed that phosphorylation of Tyr-93 located within domain 1 of NS5A, but not of any other tyrosine residue, is crucial for complex formation. In line with these findings, effective replication of subgenomic HCV replicons as well as production of infectious virus particles in mammalian cell culture models were clearly dependent on the presence of tyrosine at position 93 of NS5A. These findings indicate that phosphorylated Tyr-93 in NS5A plays an important role during viral replication by facilitating NS5A's interaction with the SH2 domain of c-Src.
Identifiants
pubmed: 30862675
pii: S0021-9258(20)36814-9
doi: 10.1074/jbc.RA119.007656
pmc: PMC6509489
doi:
Substances chimiques
MAS1 protein, human
0
Proto-Oncogene Mas
0
Viral Nonstructural Proteins
0
Tyrosine
42HK56048U
src-Family Kinases
EC 2.7.10.2
NS-5 protein, hepatitis C virus
EC 2.7.7.48
Banques de données
PDB
['1ZH1', '3FQQ', '4CL1']
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
7388-7402Informations de copyright
© 2019 Klinker et al.
Références
Biochem Biophys Res Commun. 2004 Nov 26;324(4):1155-64
pubmed: 15504335
Proc Natl Acad Sci U S A. 2004 Aug 31;101(35):13038-43
pubmed: 15326295
J Mol Biol. 2012 Jul 20;420(4-5):310-23
pubmed: 22543239
Virus Res. 2007 Aug;127(2):195-207
pubmed: 17428568
J Biol Chem. 2004 Nov 19;279(47):48576-87
pubmed: 15339921
J Chem Theory Comput. 2013 Jun 11;9(6):2733-2748
pubmed: 23914143
J Virol. 2015 Mar;89(6):3123-35
pubmed: 25552726
J Virol. 2010 Jan;84(1):482-91
pubmed: 19812153
J Comput Chem. 2004 Jul 15;25(9):1157-74
pubmed: 15116359
Biol Chem. 2009 Oct;390(10):1013-32
pubmed: 19642875
J Biol Chem. 2002 Mar 29;277(13):11149-55
pubmed: 11801599
Nature. 2005 May 19;435(7040):374-9
pubmed: 15902263
Cell Host Microbe. 2014 Nov 12;16(5):569-79
pubmed: 25525790
J Mol Model. 2006 Feb;12(3):281-9
pubmed: 16240095
J Pept Sci. 2014 May;20(5):334-40
pubmed: 24616074
J Chem Theory Comput. 2015 Apr 14;11(4):1864-74
pubmed: 26574392
J Cell Physiol. 2012 Mar;227(3):1090-7
pubmed: 21567396
J Virol. 2013 Oct;87(19):10612-27
pubmed: 23885072
J Chem Theory Comput. 2015 Aug 11;11(8):3696-713
pubmed: 26574453
Science. 1999 Jul 2;285(5424):110-3
pubmed: 10390360
J Virol. 2005 Mar;79(5):3187-94
pubmed: 15709040
Hepatology. 2011 Apr;53(4):1127-36
pubmed: 21480319
Cold Spring Harb Perspect Biol. 2013 Dec 01;5(12):a008987
pubmed: 24296166
mBio. 2015 Jul 07;6(4):e00759
pubmed: 26152585
Biomol NMR Assign. 2011 Oct;5(2):241-3
pubmed: 21516467
J Biol Chem. 2012 Dec 28;287(53):44249-60
pubmed: 23152499
J Virol. 2008 Feb;82(3):1073-83
pubmed: 18032500
J Virol. 2007 Dec;81(23):13168-79
pubmed: 17881454
PLoS Pathog. 2009 Jun;5(6):e1000475
pubmed: 19521536
PLoS Pathog. 2008 Mar 28;4(3):e1000035
pubmed: 18369481
J Virol. 2002 Dec;76(24):13001-14
pubmed: 12438626
Methods. 2013 Feb;59(2):233-48
pubmed: 23009812
Proc Natl Acad Sci U S A. 2006 May 9;103(19):7408-13
pubmed: 16651538
Biophys J. 2015 Oct 6;109(7):1483-96
pubmed: 26445449
Nat Cell Biol. 2007 Sep;9(9):1089-97
pubmed: 17721513
Nat Rev Microbiol. 2007 Jun;5(6):453-63
pubmed: 17487147
Proteins. 2008 Nov 15;73(3):765-83
pubmed: 18498103
J Virol. 2014 Jul;88(13):7541-55
pubmed: 24760886
Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3199-203
pubmed: 7536927
Gastroenterology. 2006 Mar;130(3):794-809
pubmed: 16530520
J Biol Chem. 2015 Sep 4;290(36):21857-64
pubmed: 26203192
Cell. 1993 Mar 12;72(5):779-90
pubmed: 7680960
J Gen Virol. 2015 Apr;96(Pt 4):727-738
pubmed: 25481754
Chem Biol. 2000 Jan;7(1):R3-8
pubmed: 10662684
Cancer Res. 1982 Sep;42(9):3858-63
pubmed: 6286115
PLoS One. 2012;7(10):e46634
pubmed: 23077515
J Magn Reson. 2012 Jan;214(1):329-34
pubmed: 22123230
Methods. 2001 Dec;25(4):402-8
pubmed: 11846609
Biochemistry. 2013 Sep 10;52(36):6160-8
pubmed: 23947833
Cell. 1993 Mar 12;72(5):767-78
pubmed: 7680959
Sci Rep. 2014 Apr 23;4:4765
pubmed: 24755925
PLoS Pathog. 2008 Mar 21;4(3):e1000032
pubmed: 18369478
J Gen Virol. 2004 Mar;85(Pt 3):721-729
pubmed: 14993658
J Biomol NMR. 2013 Apr;55(4):311-21
pubmed: 23435576
J Virol. 2009 May;83(9):4395-403
pubmed: 19244328
Science. 2000 Dec 8;290(5498):1972-4
pubmed: 11110665
J Virol. 2013 Jan;87(2):1255-60
pubmed: 23115292
Protein Sci. 2014 Jun;23(6):723-34
pubmed: 24639329
EBioMedicine. 2018 Feb;28:151-161
pubmed: 29331674
Biochemistry. 2017 Jun 20;56(24):3029-3048
pubmed: 28535337
J Virol. 2014 Feb;88(3):1421-32
pubmed: 24257600
Cell Rep. 2013 Apr 25;3(4):1293-305
pubmed: 23545499