Identification of an essential domain in the herpes simplex virus 1 UL34 protein that is necessary and sufficient to interact with UL31 protein.
Previous results have indicated that the herpes simplex virus 1 UL31 and UL34 proteins interact and form a complex at the inner nuclear membranes of infected cells, where both play important roles in the envelopment of nucleocapsids at the inner nuclear membrane. In the work described here, mapping studies using glutathione S-transferase pull-down assays indicated that amino acids 137 to 181 of the UL34 protein are sufficient to mediate an interaction with the UL31 protein. A recombinant virus (v3480) lacking UL34 codons 138 to 181 was constructed. Similar to a UL34 null virus, v3480 failed to replicate on Vero cells and grew to a limited extent on rabbit skin cells. A UL34-expressing cell line restored v3480 growth and plaque formation. Similar to the localization of UL31 protein in cells infected with a UL34 null virus, the UL31 protein was present in the nuclei of Hep2 cells infected with v3480. Hep2 cells infected with v3480 contained the UL34 protein in the cytoplasm, the nucleus, and the nuclear membrane, and this was noted to be similar to the appearance of cells infected with a UL31 null virus. In transient expression assays, the interaction between UL34 amino acids 137 to 181 and the UL31 protein was sufficiently robust to target green fluorescent protein and emerin to intranuclear sites that contained the UL31 protein. These data indicate that amino acids 137 to 181 of the UL34 protein are (i) sufficient to mediate interactions with the UL31 protein in vitro and in vivo, (ii) necessary for the colocalization of UL31 and UL34 in infected cells, and (iii) essential for normal viral replication.
Liang
Li
L
Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853, USA.
Baines
Joel D
JD
D000818:Animals; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000737:chemistry; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000737:chemistry; D017930:Genes, Reporter; D005816:Genetic Complementation Test; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D018259:Herpesvirus 1, Human; D006801:Humans; D009685:Nuclear Envelope / Q000737:chemistry; D009687:Nuclear Proteins / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D014709:Vero Cells; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; C108721:UL31 protein, Human herpesvirus 1; C403420:UL34 protein, Human herpesvirus 1; D014764:Viral Proteins; D049452:Green Fluorescent Proteins
J Virol. 2004 Aug;78(15):8026-35
15254174
J Virol. 2004 Jun;78(11):5591-600
15140956
J Gen Virol. 1968 May;2(3):357-64
4300104
J Cell Biol. 1990 Mar;110(3):569-80
2307700
J Virol. 1991 Nov;65(11):5757-64
1656069
J Virol. 1992 Jul;66(7):4295-303
1318405
J Virol. 1992 Aug;66(8):5168-74
1321297
Anal Biochem. 1993 Apr;210(1):179-87
8489015
J Virol. 1993 Nov;67(11):6348-56
7692079
J Biol Chem. 1994 Jul 1;269(26):17401-10
8021242
Nat Genet. 1994 Dec;8(4):323-7
7894480
J Virol. 1995 Sep;69(9):5401-13
7636985
J Cell Biol. 1995 Oct;131(1):33-44
7559784
J Virol. 1997 Jul;71(7):5012-24
9188566
J Virol. 1997 Nov;71(11):8307-15
9343183
J Virol. 1999 May;73(5):4110-9
10196307
J Cell Sci. 1999 Jun;112 ( Pt 11):1709-19
10318763
J Virol. 1999 Oct;73(10):8503-11
10482603
Int J Biochem Cell Biol. 1999 Sep;31(9):891-4
10533281
J Virol. 2000 Jan;74(1):117-29
10590098
J Virol. 2000 Feb;74(3):1355-63
10627546
J Virol. 2000 Feb;74(4):1827-39
10644356
J Gen Virol. 2000 Oct;81(Pt 10):2397-405
10993927
Proc Natl Acad Sci U S A. 2000 Sep 26;97(20):11002-7
11005871
J Virol. 2000 Nov;74(21):10063-73
11024135
J Virol. 2001 Apr;75(8):3675-84
11264357
Genomics. 2001 Apr 1;73(1):56-65
11352566
J Virol. 2001 Jun;75(12):5697-702
11356979
J Gen Virol. 2001 Jun;82(Pt 6):1423-8
11369887
J Virol. 2001 Sep;75(18):8803-17
11507225
J Virol. 2001 Dec;75(24):12209-19
11711612
J Virol. 2002 Jan;76(1):364-78
11739701
J Virol. 2002 Feb;76(4):1537-47
11799148
J Virol. 2002 Sep;76(17):8939-52
12163613
Virology. 2002 Sep 1;300(2):189-204
12350350
J Virol. 2003 Jan;77(2):1382-91
12502854
Biochemistry. 2003 May 6;42(17):4819-28
12718522
J Virol. 2003 Jul;77(13):7601-10
12805460
J Virol. 2004 Jan;78(1):399-412
14671121
Curr Opin Cell Biol. 2004 Feb;16(1):73-9
15037308
J Virol. 2004 Jun;78(11):5564-75
15140953
Virology. 2004 Nov 10;329(1):68-76
15476875