Identification and subcellular localization of porcine deltacoronavirus accessory protein NS6.
Porcine deltacoronavirus (PDCoV) is an emerging swine enteric coronavirus. Accessory proteins are genus-specific for coronavirus, and two putative accessory proteins, NS6 and NS7, are predicted to be encoded by PDCoV; however, this remains to be confirmed experimentally. Here, we identified the leader-body junction sites of NS6 subgenomic RNA (sgRNA) and found that the actual transcription regulatory sequence (TRS) utilized by NS6 is non-canonical and is located upstream of the predicted TRS. Using the purified NS6 from an Escherichia coli expression system, we obtained two anti-NS6 monoclonal antibodies that could detect the predicted NS6 in cells infected with PDCoV or transfected with NS6-expressing plasmids. Further studies revealed that NS6 is always localized in the cytoplasm of PDCoV-infected cells, mainly co-localizing with the endoplasmic reticulum (ER) and ER-Golgi intermediate compartments, as well as partially with the Golgi apparatus. Together, our results identify the NS6 sgRNA and demonstrate its expression in PDCoV-infected cells.
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
Puxian
P
Fang
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
Liurong
L
Fang
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
Xiaorong
X
Liu
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
Yingying
Y
Hong
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
Yongle
Y
Wang
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
Nan
N
Dong
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
Panpan
P
Ma
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; Department of Immunology and Aetology, College of Basic Medicine, Hubei University of Chinese Medicine, Wuhan 430065, China.
Jing
J
Bi
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China.
Dang
D
Wang
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China. Electronic address: vet@mail.hzau.edu.cn.
Shaobo
S
Xiao
D000818:Animals; D001483:Base Sequence; D017934:Coronavirus / Q000502:physiology*; D015870:Gene Expression; D016679:Genome, Viral; D042541:Intracellular Space / Q000378:metabolism; D016366:Open Reading Frames; D021381:Protein Transport; D012367:RNA, Viral; D011994:Recombinant Proteins; D013552:Swine; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D012367:RNA, Viral; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins
Accessory protein; NS6; Porcine deltacoronavirus; Subcelluar localization; Subgenomic RNA
J Virol. 2004 Jan;78(2):980-94
14694129
Virus Res. 2016 Dec 2;226:71-84
27270129
Transbound Emerg Dis. 2016 Jun;63(3):248-52
26968326
Genome Announc. 2014 Apr 10;2(2):
24723704
J Clin Microbiol. 2015 May;53(5):1537-48
25740769
Virus Res. 2016 Dec 2;226:50-59
27086031
Proc Natl Acad Sci U S A. 1999 Oct 12;96(21):12056-61
10518575
Emerg Infect Dis. 2014 Jul;20(7):1227-30
24964136
J Biotechnol. 2009 Apr 20;141(1-2):91-6
19428736
J Virol. 2005 May;79(9):5288-95
15827143
Viruses. 2012 Nov 07;4(11):2902-23
23202509
J Virol. 2005 Dec;79(23):14909-22
16282490
Virology. 2007 Dec 5;369(1):92-104
17706262
Antiviral Res. 2014 Sep;109:97-109
24995382
Arch Virol. 2016 Oct;161(10):2909-11
27424024
Virology. 2002 Apr 25;296(1):177-89
12036329
Viruses. 2010 Aug;2(8):1804-20
21994708
Virology. 2016 Aug;495:10-7
27152478
J Gen Virol. 2001 Feb;82(Pt 2):385-396
11161278
J Virol. 2013 Feb;87(4):2128-36
23221558
Emerg Infect Dis. 2015 Dec;21(12):2254-5
26584185
J Virol. 2012 Apr;86(7):3995-4008
22278237
Genome Announc. 2015 Sep 03;3(5):
26337879
J Virol. 2013 Nov;87(22):12489-95
24027320
J Virol. 2009 Mar;83(5):2368-73
19091867
J Virol. 2009 Jul;83(13):6631-40
19403678
mBio. 2016 Mar 29;7(2):e00258
27025250
J Virol. 1994 Oct;68(10):6523-34
8083990
J Gen Virol. 2005 Jul;86(Pt 7):1921-1930
15958670
J Virol. 2006 Aug;80(15):7287-94
16840309
J Gen Virol. 2005 Aug;86(Pt 8):2249-2253
16033972
Genome Announc. 2016 May 26;4(3):
27231358
Virus Res. 2015 Oct 2;208:136-45
26103099
J Virol. 2007 Jan;81(2):548-57
17108024
J Virol. 2005 Jul;79(13):8065-78
15956552
J Virol. 2004 Apr;78(8):3863-71
15047802
J Virol. 2016 Jun 24;90(14):6573-6582
27170748
Emerg Infect Dis. 2016 Apr;22(4):757-9
26982324
Genome Announc. 2014 Nov 26;2(6):
25428966
J Virol. 2007 Jan;81(1):20-9
16928755
J Virol. 2009 Jan;83(2):908-17
18971277
Emerg Infect Dis. 2015 Apr;21(4):650-4
25811229
Genome Announc. 2014 Apr 10;2(2):
24723718
mBio. 2015 Mar 10;6(2):e00064
25759498
Antiviral Res. 2006 Nov;72(2):78-88
16820226
Transbound Emerg Dis. 2015 Dec;62(6):575-80
26250097
J Virol. 2004 Jul;78(14):7311-8
15220404
Emerg Infect Dis. 2014 Sep;20(9):1594-5
25153521
Emerg Infect Dis. 2014 Aug;20(8):1347-50
25075556
J Virol. 2014 May;88(9):4866-76
24522921
Genome Announc. 2015 Oct 29;3(5):
26514769
Virology. 2015 Aug;482:51-9
25817405