Identification and characterization of the ICP22 protein of equine herpesvirus 1

V. Roger Holden, Gretchen B Caughman, Yuhe Zhao, Ronald N. Harty, Dennis J. O'Callaghan

Research output: Contribution to journalArticle

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Abstract

The equine herpesvirus 1 (EHV-1) homolog of herpes simplex virus type 1 ICP22 is differently expressed from the fourth open reading frame of the inverted repeat (IR4) as a 1.4-kb early mRNA and a 1.7-kb late mRNA which are 3' coterminal (V. R. Holden, R. R. Yalamanchili, R. N. Harty, and D. J. O'Callaghan, J. Virol. 66:664-673, 1992). To extend the characterization of IR4 at the protein level, the synthesis and intracellular localization of the IR4 protein were investigated. Antiserum raised against either a synthetic peptide corresponding to amino acids 270 to 286 or against a TrpE-IR4 fusion protein (IR4 residues 13 to 150) was used to identify the IR4 protein. Western immunoblot analysis revealed that IR4 is expressed abundantly from an open reading frame composed of 293 codons as a family of proteins that migrate between 42 to 47 kDa. The intracellular localization of IR4 was examined by cell fractionation, indirect immunofluorescence, and laser- scanning confocal microscopy. These studies revealed that IR4 is localized predominantly in the nucleus and is dispersed uniformly throughout the nucleus. Interestingly, when IR4 is expressed transiently in COS-1 or LTK- cells, a punctate staining pattern within the nucleus is observed by indirect immunofluorescence. Cells transfected with an IR4 mutant construct that encodes a C-terminal truncated (19 amino acids) IR4 protein exhibited greatly reduced intranuclear accumulation of the IR4 protein, indicating that this domain possesses an important intranuclear localization signal. Western blot analysis of EHV-1 virion proteins revealed that IR4 proteins are structural components of the virions. Surprisingly, the 42-kDa species, which is the least abundant and the least modified form of the IR4 protein family in infected cell extracts, was the most abundant IR4 protein present in purified virions.

Original languageEnglish (US)
Pages (from-to)4329-4340
Number of pages12
JournalJournal of Virology
Volume68
Issue number7
StatePublished - Jul 1 1994

Fingerprint

Equid Herpesvirus 1
Equid herpesvirus 1
Proteins
proteins
Virion
virion
Indirect Fluorescent Antibody Technique
Open Reading Frames
fluorescent antibody technique
open reading frames
Western Blotting
Cell Fractionation
Amino Acids
cell fractionation
Messenger RNA
Human herpesvirus 1
amino acids
Human Herpesvirus 1
synthetic peptides
Cell Extracts

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Holden, V. R., Caughman, G. B., Zhao, Y., Harty, R. N., & O'Callaghan, D. J. (1994). Identification and characterization of the ICP22 protein of equine herpesvirus 1. Journal of Virology, 68(7), 4329-4340.

Identification and characterization of the ICP22 protein of equine herpesvirus 1. / Holden, V. Roger; Caughman, Gretchen B; Zhao, Yuhe; Harty, Ronald N.; O'Callaghan, Dennis J.

In: Journal of Virology, Vol. 68, No. 7, 01.07.1994, p. 4329-4340.

Research output: Contribution to journalArticle

Holden, VR, Caughman, GB, Zhao, Y, Harty, RN & O'Callaghan, DJ 1994, 'Identification and characterization of the ICP22 protein of equine herpesvirus 1', Journal of Virology, vol. 68, no. 7, pp. 4329-4340.
Holden VR, Caughman GB, Zhao Y, Harty RN, O'Callaghan DJ. Identification and characterization of the ICP22 protein of equine herpesvirus 1. Journal of Virology. 1994 Jul 1;68(7):4329-4340.
Holden, V. Roger ; Caughman, Gretchen B ; Zhao, Yuhe ; Harty, Ronald N. ; O'Callaghan, Dennis J. / Identification and characterization of the ICP22 protein of equine herpesvirus 1. In: Journal of Virology. 1994 ; Vol. 68, No. 7. pp. 4329-4340.
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