Identification and characterization of the host protein DNAJC14 as a broadly active flavivirus replication modulator.
Viruses in the Flavivirus genus of the Flaviviridae family are arthropod-transmitted and contribute to staggering numbers of human infections and significant deaths annually across the globe. To identify cellular factors with antiviral activity against flaviviruses, we screened a cDNA library using an iterative approach. We identified a mammalian Hsp40 chaperone protein (DNAJC14) that when overexpressed was able to mediate protection from yellow fever virus (YFV)-induced cell death. Further studies revealed that DNAJC14 inhibits YFV at the step of viral RNA replication. Since replication of bovine viral diarrhea virus (BVDV), a member of the related Pestivirus genus, is also known to be modulated by DNAJC14, we tested the effect of this host factor on diverse Flaviviridae family members. Flaviviruses, including the pathogenic Asibi strain of YFV, Kunjin, and tick-borne Langat virus, as well as a Hepacivirus, hepatitis C virus (HCV), all were inhibited by overexpression of DNAJC14. Mutagenesis showed that both the J-domain and the C-terminal domain, which mediates self-interaction, are required for anti-YFV activity. We found that DNAJC14 does not block YFV nor HCV NS2-3 cleavage, and using non-inhibitory mutants demonstrate that DNAJC14 is recruited to YFV replication complexes. Immunofluorescence analysis demonstrated that endogenous DNAJC14 rearranges during infection and is found in replication complexes identified by dsRNA staining. Interestingly, silencing of endogenous DNAJC14 results in impaired YFV replication suggesting a requirement for DNAJC14 in YFV replication complex assembly. Finally, the antiviral activity of overexpressed DNAJC14 occurs in a time- and dose-dependent manner. DNAJC14 overexpression may disrupt the proper stoichiometry resulting in inhibition, which can be overcome upon restoration of the optimal ratios due to the accumulation of viral nonstructural proteins. Our findings, together with previously published work, suggest that the members of the Flaviviridae family have evolved in unique and important ways to interact with this host Hsp40 chaperone molecule.
Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, New York, USA.
Zhigang
Z
Yi
Lindsey
L
Sperzel
Cindy
C
Nürnberger
Peter J
PJ
Bredenbeek
Kirk J
KJ
Lubick
Sonja M
SM
Best
Cristina T
CT
Stoyanov
Lok Man J
LM
Law
Zhenghong
Z
Yuan
Charles M
CM
Rice
Margaret R
MR
MacDonald
D000818:Animals; D002417:Cattle; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D006224:Cricetinae; D005326:Fetal Proteins / Q000276:immunology* / Q000378:metabolism; D054884:Host-Pathogen Interactions / Q000276:immunology*; D006801:Humans; D018832:Molecular Chaperones / Q000276:immunology* / Q000378:metabolism; D014709:Vero Cells; D014779:Virus Replication / Q000276:immunology*; D015004:Yellow Fever / Q000276:immunology* / Q000821:virology; D015005:Yellow fever virus / Q000276:immunology* / Q000472:pathogenicity
C476535:DNAJC14 protein, human; D005326:Fetal Proteins; D018832:Molecular Chaperones
Vaccine. 2010 Aug 9;28(35):5676-85
20600494
Nature. 2006 Aug 17;442(7104):831-5
16862121
Nat Struct Mol Biol. 2006 Oct;13(10):865-70
17021621
J Virol. 1998 Sep;72(9):7270-9
9696822
J Virol. 2007 Dec;81(24):13509-18
17928353
Virology. 1999 Nov 10;264(1):66-75
10544130
Nat Med. 2004 Dec;10(12 Suppl):S98-109
15577938
Trends Biochem Sci. 1998 Jun;23(6):222-7
9644977
Nature. 2007 Apr 12;446(7137):801-5
17325668
Proc Natl Acad Sci U S A. 2006 Jan 31;103(5):1510-5
16432213
Mol Cell Proteomics. 2005 Dec;4(12):1933-41
16155292
J Virol. 2003 Oct;77(19):10663-9
12970452
J Virol. 2000 Sep;74(18):8358-67
10954535
J Virol. 2004 Mar;78(5):2414-25
14963137
J Virol. 2002 May;76(10):4773-84
11967294
J Virol. 2004 Oct;78(19):10765-75
15367643
Cell. 1998 Feb 6;92(3):351-66
9476895
Cell. 2000 Apr 14;101(2):173-85
10786833
J Virol. 1991 Nov;65(11):6042-50
1833562
Antiviral Res. 2009 Jan;81(1):6-15
18796313
J Virol. 2003 Nov;77(21):11555-62
14557641
J Virol. 2004 Dec;78(23):12781-7
15542630
Biochemistry. 2008 Jul 8;47(27):7001-11
18557634
Nat Rev Mol Cell Biol. 2010 Aug;11(8):579-92
20651708
Virology. 2008 Nov 25;381(2):268-76
18823640
J Virol. 1997 Dec;71(12):9608-17
9371625
Vaccine. 2010 Jun 23;28(29):4644-52
20451637
Virus Res. 2001 Nov 5;79(1-2):59-69
11551646
Proc Natl Acad Sci U S A. 1990 Nov;87(22):8898-902
2147282
J Virol. 1999 Oct;73(10):8083-94
10482557
Hepatology. 2008 Dec;48(6):1843-50
19003912
Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1746-51
9465088
J Biol Chem. 2005 Aug 19;280(33):29604-11
15980068
J Virol. 2002 Dec;76(24):13001-14
12438626
J Virol. 2005 Aug;79(15):9746-55
16014936
J Virol. 2010 May;84(9):4504-12
20181706
J Virol. 2003 Jan;77(2):1644-8
12502883
J Virol. 2001 Oct;75(19):9470-82
11533209
Nat Cell Biol. 2001 May;3(5):492-8
11331877
Protein Pept Lett. 2009;16(6):606-12
19519518
Virology. 1990 Jul;177(1):159-74
2353452
Lancet. 2008 Feb 9;371(9611):500-9
18262042
J Gen Virol. 2003 May;84(Pt 5):1261-1268
12692292
Proc Natl Acad Sci U S A. 2002 Sep 3;99(18):11914-9
12154227
Virology. 2005 Jan 20;331(2):247-59
15629769
Virology. 1983 Feb;125(1):8-17
6187129
Lab Invest. 2004 May;84(5):658-61
15105816
Nat Biotechnol. 2010 Feb;28(2):167-71
20118917
J Virol. 2010 Apr;84(7):3503-15
20106931
J Virol. 2010 Jul;84(13):6720-32
20392851