Interaction of the herpes simplex virus type 1 packaging protein UL15 with full-length and deleted forms of the UL28 protein

被引:39
作者
Abbotts, AP [1 ]
Preston, VG [1 ]
Hughes, M [1 ]
Patel, AH [1 ]
Stow, ND [1 ]
机构
[1] Univ Glasgow, Inst Virol, MRC, Virol Unit, Glasgow G11 5JR, Lanark, Scotland
关键词
D O I
10.1099/0022-1317-81-12-2999
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The UL15 and UL28 proteins of herpes simplex virus type 1 are both required for the packaging of replicated viral DNA into the viral capsid. We have expressed UL28 and a functional epitope-tagged form of UL15 in mammalian and insect cells. Immunoprecipitation experiments confirmed that the two proteins can interact. In agreement with previous results, UL15, when expressed alone, entered the nucleus but UL28 remained cytoplasmic. When co-expressed the two proteins co-localized in the nucleus. Six UL28 deletion mutants were constructed and similarly analysed. The results obtained by immunoprecipitation and immunofluorescence were consistent and demonstrate that at least two separate regions of the UL28 polypeptide chain have the ability to interact with UL15. Surprisingly, three of the mutants prevented the UL15 protein from localizing to the cell nucleus, and these were not functional in a transient DNA packaging assay. Of the three UL28 mutant proteins that entered the nucleus with UL15, one containing an internal deletion of 13 amino acids was able to complement a UL28 null mutant in both DNA packaging and virus yield assays, demonstrating that this region of the protein is not essential for function. In addition to interacting with the UL28 protein we also demonstrated that UL15 molecules can interact with each other, and that sequences within the second exon contribute to this interaction.
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页码:2999 / 3009
页数:11
相关论文
共 46 条
[1]   CHARACTERIZATION OF A HERPES-SIMPLEX VIRUS TYPE-1 MUTANT WHICH HAS A TEMPERATURE-SENSITIVE DEFECT IN PENETRATION OF CELLS AND ASSEMBLY OF CAPSIDS [J].
ADDISON, C ;
RIXON, FJ ;
PALFREYMAN, JW ;
OHARA, M ;
PRESTON, VG .
VIROLOGY, 1984, 138 (02) :246-259
[2]   HERPES-SIMPLEX VIRUS TYPE-1 UL28 GENE-PRODUCT IS IMPORTANT FOR THE FORMATION OF MATURE CAPSIDS [J].
ADDISON, C ;
RIXON, FJ ;
PRESTON, VG .
JOURNAL OF GENERAL VIROLOGY, 1990, 71 :2377-2384
[3]   Characterization of an essential HSV-1 protein encoded by the UL25 gene reported to be involved in virus penetration and capsid assembly [J].
Ali, MA ;
Forghani, B ;
Cantin, EM .
VIROLOGY, 1996, 216 (01) :278-283
[4]   The U(L)15 gene of herpes simplex virus type 1 contains within its second exon a novel open reading frame that is translated in frame with the U(L)15 gene product [J].
Baines, JD ;
Cunningham, C ;
Nalwanga, D ;
Davison, A .
JOURNAL OF VIROLOGY, 1997, 71 (04) :2666-2673
[5]  
BISHOP DHL, 1992, SEMINARS VIROLOGY, V3, P253
[6]   The gene product of human cytomegalovirus open reading frame UL56 binds the pac motif and has specific nuclease activity [J].
Bogner, E ;
Radsak, K ;
Stinski, MF .
JOURNAL OF VIROLOGY, 1998, 72 (03) :2259-2264
[7]   The terminase enzyme from bacteriophage lambda: a DNA-packaging machine [J].
Catalano, CE .
CELLULAR AND MOLECULAR LIFE SCIENCES, 2000, 57 (01) :128-148
[8]   VIRUS-DNA PACKAGING - THE STRATEGY USED BY PHAGE-LAMBDA [J].
CATALANO, CE ;
CUE, D ;
FEISS, M .
MOLECULAR MICROBIOLOGY, 1995, 16 (06) :1075-1086
[9]   CHANNEL CATFISH VIRUS - A NEW TYPE OF HERPESVIRUS [J].
DAVISON, AJ .
VIROLOGY, 1992, 186 (01) :9-14
[10]   SEQUENCE-ANALYSIS OF THE SPLICE JUNCTION IN THE TRANSCRIPT OF HERPES-SIMPLEX VIRUS TYPE-1 GENE UL15 [J].
DOLAN, A ;
ARBUCKLE, M ;
MCGEOCH, DJ .
VIRUS RESEARCH, 1991, 20 (01) :97-104