Structural Description of the Nipah Virus Phosphoprotein and Its Interaction with STAT1.
Journal
Biophysical journal
ISSN: 1542-0086
Titre abrégé: Biophys J
Pays: United States
ID NLM: 0370626
Informations de publication
Date de publication:
19 05 2020
19 05 2020
Historique:
received:
18
07
2019
revised:
31
03
2020
accepted:
06
04
2020
pubmed:
30
4
2020
medline:
15
5
2021
entrez:
30
4
2020
Statut:
ppublish
Résumé
The structural characterization of modular proteins containing long intrinsically disordered regions intercalated with folded domains is complicated by their conformational diversity and flexibility and requires the integration of multiple experimental approaches. Nipah virus (NiV) phosphoprotein, an essential component of the viral RNA transcription/replication machine and a component of the viral arsenal that hijacks cellular components and counteracts host immune responses, is a prototypical model for such modular proteins. Curiously, the phosphoprotein of NiV is significantly longer than the corresponding protein of other paramyxoviruses. Here, we combine multiple biophysical methods, including x-ray crystallography, NMR spectroscopy, and small angle x-ray scattering, to characterize the structure of this protein and provide an atomistic representation of the full-length protein in the form of a conformational ensemble. We show that full-length NiV phosphoprotein is tetrameric, and we solve the crystal structure of its tetramerization domain. Using NMR spectroscopy and small angle x-ray scattering, we show that the long N-terminal intrinsically disordered region and the linker connecting the tetramerization domain to the C-terminal X domain exchange between multiple conformations while containing short regions of residual secondary structure. Some of these transient helices are known to interact with partners, whereas others represent putative binding sites for yet unidentified proteins. Finally, using NMR spectroscopy and isothermal titration calorimetry, we map a region of the phosphoprotein, comprising residues between 110 and 140 and common to the V and W proteins, that binds with weak affinity to STAT1 and confirm the involvement of key amino acids of the viral protein in this interaction. This provides new, to our knowledge, insights into how the phosphoprotein and the nonstructural V and W proteins of NiV perform their multiple functions.
Identifiants
pubmed: 32348724
pii: S0006-3495(20)30328-3
doi: 10.1016/j.bpj.2020.04.010
pmc: PMC7231922
pii:
doi:
Substances chimiques
Phosphoproteins
0
Viral Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2470-2488Informations de copyright
Copyright © 2020 Biophysical Society. Published by Elsevier Inc. All rights reserved.
Références
J Gen Virol. 2006 Dec;87(Pt 12):3649-3653
pubmed: 17098981
J Mol Biol. 2004 May 14;338(5):1015-26
pubmed: 15111064
J Virol. 2003 Nov;77(21):11842-5
pubmed: 14557668
Protein Sci. 2006 Dec;15(12):2795-804
pubmed: 17088319
Protein Sci. 2011 Mar;20(3):542-56
pubmed: 21207454
Sci Adv. 2018 Aug 22;4(8):eaat7778
pubmed: 30140745
Nat Commun. 2018 Sep 12;9(1):3703
pubmed: 30209309
Biochemistry. 1995 May 23;34(20):6784-94
pubmed: 7756310
J Am Chem Soc. 2010 Feb 3;132(4):1270-2
pubmed: 20063887
Proc Natl Acad Sci U S A. 2020 Jan 28;117(4):2099-2107
pubmed: 31953264
J Biol Chem. 2011 Apr 15;286(15):13583-602
pubmed: 21317293
Infect Genet Evol. 2010 Jan;10(1):97-107
pubmed: 19900582
Cell Rep. 2020 Jan 7;30(1):53-60.e5
pubmed: 31914397
J Virol. 2014 Jan;88(1):758-62
pubmed: 24155387
Adv Virus Res. 2017;98:1-55
pubmed: 28433050
J Comput Chem. 2004 Oct;25(13):1605-12
pubmed: 15264254
Angew Chem Int Ed Engl. 2016 Aug 1;55(32):9356-60
pubmed: 27270664
J Infect Dis. 2019 May 24;219(12):1867-1878
pubmed: 30364984
Biophys J. 2009 Nov 18;97(10):2839-45
pubmed: 19917239
J Mol Biol. 1993 Dec 5;234(3):779-815
pubmed: 8254673
J Gen Virol. 2004 Mar;85(Pt 3):701-707
pubmed: 14993656
Proteins. 2006 Oct 1;65(1):1-14
pubmed: 16856179
Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21
pubmed: 20057044
J Virol. 2000 Nov;74(21):9972-9
pubmed: 11024125
Adv Exp Med Biol. 2017;1009:47-64
pubmed: 29218553
Nat Struct Mol Biol. 2014 Sep;21(9):754-9
pubmed: 25108352
J Virol. 2013 Sep;87(17):9569-78
pubmed: 23785215
PLoS One. 2010 Jul 21;5(7):e11684
pubmed: 20657787
J Virol. 2009 Aug;83(16):7828-41
pubmed: 19515782
J Gen Virol. 2003 Dec;84(Pt 12):3239-3252
pubmed: 14645906
Science. 2015 May 8;348(6235):704-7
pubmed: 25883315
Acta Crystallogr D Biol Crystallogr. 2008 Jan;64(Pt 1):61-9
pubmed: 18094468
Sci Adv. 2019 May 08;5(5):eaaw3702
pubmed: 31086822
J Biol Chem. 2003 Nov 7;278(45):44567-73
pubmed: 12944395
J Virol. 2008 Sep;82(17):8330-8
pubmed: 18579593
Biochemistry. 2007 Sep 11;46(36):10328-38
pubmed: 17705401
Cell. 1998 May 29;93(5):827-39
pubmed: 9630226
PLoS Pathog. 2016 Dec 9;12(12):e1006058
pubmed: 27936158
J Virol. 2009 Apr;83(8):3982-7
pubmed: 19211754
Sci Rep. 2018 Jan 10;8(1):358
pubmed: 29321677
PLoS Pathog. 2013;9(9):e1003631
pubmed: 24086133
PLoS Pathog. 2011 Sep;7(9):e1002248
pubmed: 21960769
J Mol Biol. 1999 Oct 22;293(2):321-31
pubmed: 10550212
Cell. 2019 Sep 19;179(1):193-204.e14
pubmed: 31495574
Virology. 2015 Mar;477:42-55
pubmed: 25637789
Structure. 2019 Apr 2;27(4):660-668.e4
pubmed: 30799076
Future Microbiol. 2010 Jan;5(1):9-13
pubmed: 20020826
J Virol. 2013 Jul;87(13):7558-68
pubmed: 23637399
J Biomol NMR. 1995 Nov;6(3):277-93
pubmed: 8520220
Biochemistry. 1993 Dec 7;32(48):13288-98
pubmed: 8241185
PLoS Pathog. 2018 Feb 9;14(2):e1006889
pubmed: 29425244
Acta Crystallogr D Biol Crystallogr. 2014 Jun;70(Pt 6):1589-603
pubmed: 24914970
J Am Chem Soc. 2009 Dec 16;131(49):17908-18
pubmed: 19908838
Science. 2000 May 26;288(5470):1432-5
pubmed: 10827955
J Virol. 2004 Jun;78(11):5633-41
pubmed: 15140960
J Virol. 2010 Oct;84(19):9793-9
pubmed: 20668089
Cell Mol Life Sci. 2017 Sep;74(17):3069-3090
pubmed: 28589442
J Virol. 2010 Apr;84(7):3707-10
pubmed: 20089657
J Virol. 2004 May;78(10):5358-67
pubmed: 15113915
Acta Crystallogr D Biol Crystallogr. 2012 Apr;68(Pt 4):352-67
pubmed: 22505256
Antiviral Res. 2013 Oct;100(1):38-43
pubmed: 23911335
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
pubmed: 15572765
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
pubmed: 20124702
EMBO J. 1996 Oct 15;15(20):5616-26
pubmed: 8896455
Protein Pept Lett. 2016;23(3):217-31
pubmed: 26732245
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):125-32
pubmed: 20124692
Sci Adv. 2017 Feb 03;3(2):e1602350
pubmed: 28168220
J Virol. 2002 Nov;76(22):11476-83
pubmed: 12388709
J Gen Virol. 2004 Jun;85(Pt 6):1675-1684
pubmed: 15166452
Virus Res. 2003 Mar;92(1):55-65
pubmed: 12606076
Mol Biosyst. 2012 Jan;8(1):151-67
pubmed: 21947276
J Mol Biol. 2009 May 22;388(5):978-96
pubmed: 19341745
World J Virol. 2013 May 12;2(2):57-70
pubmed: 24175230
J Magn Reson. 2007 Jul;187(1):163-9
pubmed: 17468025
EMBO J. 2009 Apr 8;28(7):948-58
pubmed: 19214187
J Virol. 2008 Jul;82(13):6259-71
pubmed: 18417573
J Comput Chem. 2004 Apr 15;25(5):712-24
pubmed: 14978714
Nature. 2020 Jan;577(7789):275-279
pubmed: 31698413
Proc Natl Acad Sci U S A. 2004 Aug 24;101(34):12497-502
pubmed: 15314216
Virology. 2004 Feb 20;319(2):201-11
pubmed: 14980481
J Virol. 2013 Jun;87(12):7166-9
pubmed: 23576502
Proc Natl Acad Sci U S A. 2005 Nov 22;102(47):17002-7
pubmed: 16284250
Nat Struct Biol. 2000 Sep;7(9):777-81
pubmed: 10966649
Cell Mol Life Sci. 1997 Aug;53(8):621-45
pubmed: 9351466
Adv Virus Res. 2017;97:143-185
pubmed: 28057258
PLoS One. 2012;7(3):e31719
pubmed: 22403617
Trends Biochem Sci. 2011 Mar;36(3):159-69
pubmed: 21146412
Front Microbiol. 2019 Jul 10;10:1490
pubmed: 31354644
Sci Rep. 2018 Oct 30;8(1):15994
pubmed: 30375468
J Biomol NMR. 2007 Aug;38(4):289-302
pubmed: 17610132
J Colloid Interface Sci. 1998 Jan 1;197(1):9-20
pubmed: 9466838
Biochim Biophys Acta. 1993 Feb 13;1161(2-3):244-8
pubmed: 8431473
PLoS One. 2014 Feb 25;9(2):e90003
pubmed: 24587180
J Mol Biol. 2008 Oct 3;382(2):525-38
pubmed: 18657547
J Biomol NMR. 2004 Sep;30(1):11-23
pubmed: 15452431
J Virol. 1998 Feb;72(2):1482-90
pubmed: 9445051
PLoS Pathog. 2012;8(8):e1002836
pubmed: 22879820
J Virol. 2018 Feb 12;92(5):
pubmed: 29237836
EMBO J. 2012 Mar 7;31(5):1320-9
pubmed: 22246179