title: Identification of an essential domain in the herpes simplex virus 1 UL34 protein that is necessary and sufficient to interact with UL31 protein.
abstract: 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.
authors.affiliation: Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Liang
authors.forename: Joel D
authors.initials: JD
authors.lastname: Baines
mesh_terms: 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*
chemical_list: 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
references.citation: J Virol. 2004 Aug;78(15):8026-35
references.pmid: 15254174
references.citation: J Virol. 2004 Jun;78(11):5591-600
references.pmid: 15140956
references.citation: J Gen Virol. 1968 May;2(3):357-64
references.pmid: 4300104
references.citation: J Cell Biol. 1990 Mar;110(3):569-80
references.pmid: 2307700
references.citation: J Virol. 1991 Nov;65(11):5757-64
references.pmid: 1656069
references.citation: J Virol. 1992 Jul;66(7):4295-303
references.pmid: 1318405
references.citation: J Virol. 1992 Aug;66(8):5168-74
references.pmid: 1321297
references.citation: Anal Biochem. 1993 Apr;210(1):179-87
references.pmid: 8489015
references.citation: J Virol. 1993 Nov;67(11):6348-56
references.pmid: 7692079
references.citation: J Biol Chem. 1994 Jul 1;269(26):17401-10
references.pmid: 8021242
references.citation: Nat Genet. 1994 Dec;8(4):323-7
references.pmid: 7894480
references.citation: J Virol. 1995 Sep;69(9):5401-13
references.pmid: 7636985
references.citation: J Cell Biol. 1995 Oct;131(1):33-44
references.pmid: 7559784
references.citation: J Virol. 1997 Jul;71(7):5012-24
references.pmid: 9188566
references.citation: J Virol. 1997 Nov;71(11):8307-15
references.pmid: 9343183
references.citation: J Virol. 1999 May;73(5):4110-9
references.pmid: 10196307
references.citation: J Cell Sci. 1999 Jun;112 ( Pt 11):1709-19
references.pmid: 10318763
references.citation: J Virol. 1999 Oct;73(10):8503-11
references.pmid: 10482603
references.citation: Int J Biochem Cell Biol. 1999 Sep;31(9):891-4
references.pmid: 10533281
references.citation: J Virol. 2000 Jan;74(1):117-29
references.pmid: 10590098
references.citation: J Virol. 2000 Feb;74(3):1355-63
references.pmid: 10627546
references.citation: J Virol. 2000 Feb;74(4):1827-39
references.pmid: 10644356
references.citation: J Gen Virol. 2000 Oct;81(Pt 10):2397-405
references.pmid: 10993927
references.citation: Proc Natl Acad Sci U S A. 2000 Sep 26;97(20):11002-7
references.pmid: 11005871
references.citation: J Virol. 2000 Nov;74(21):10063-73
references.pmid: 11024135
references.citation: J Virol. 2001 Apr;75(8):3675-84
references.pmid: 11264357
references.citation: Genomics. 2001 Apr 1;73(1):56-65
references.pmid: 11352566
references.citation: J Virol. 2001 Jun;75(12):5697-702
references.pmid: 11356979
references.citation: J Gen Virol. 2001 Jun;82(Pt 6):1423-8
references.pmid: 11369887
references.citation: J Virol. 2001 Sep;75(18):8803-17
references.pmid: 11507225
references.citation: J Virol. 2001 Dec;75(24):12209-19
references.pmid: 11711612
references.citation: J Virol. 2002 Jan;76(1):364-78
references.pmid: 11739701
references.citation: J Virol. 2002 Feb;76(4):1537-47
references.pmid: 11799148
references.citation: J Virol. 2002 Sep;76(17):8939-52
references.pmid: 12163613
references.citation: Virology. 2002 Sep 1;300(2):189-204
references.pmid: 12350350
references.citation: J Virol. 2003 Jan;77(2):1382-91
references.pmid: 12502854
references.citation: Biochemistry. 2003 May 6;42(17):4819-28
references.pmid: 12718522
references.citation: J Virol. 2003 Jul;77(13):7601-10
references.pmid: 12805460
references.citation: J Virol. 2004 Jan;78(1):399-412
references.pmid: 14671121
references.citation: Curr Opin Cell Biol. 2004 Feb;16(1):73-9
references.pmid: 15037308
references.citation: J Virol. 2004 Jun;78(11):5564-75
references.pmid: 15140953
references.citation: Virology. 2004 Nov 10;329(1):68-76
references.pmid: 15476875