Subcellular localization of herpes simplex virus type 1 UL51 protein and role of palmitoylation in golgi apparatus targeting

Naoki Nozawa, Tohru Daikoku, Tetsuo Koshizuka, Yohei Yamauchi, Tetsushi Yoshikawa, Yukihiro Nishiyama

Research output: Contribution to journalArticle

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Abstract

The herpes simplex virus type I (HSV-1) UL51 gene products are virion-associated phosphoproteins with apparent molecular masses of 27, 29, and 30 kDa in HSV-1-infected cells. In this study, we have investigated the intracellular localization and distribution of UL51 protein both in infected cells and in transfected cells expressing only UL51. We found that this protein colocalized closely with Golgi marker proteins such as the Golgi-58K protein and GM130 in transfected cells expressing only UL51. However, in infected cells, the UL51 protein localized to the juxtanuclear region but only partially colocalized with the Golgi maker proteins. Mutant protein analysis revealed that the N-terminal 15 amino acid residues of the UL51 protein sufficed for this Golgi localization property. The UL51 protein redistributed on addition of brefeldin A. This was prevented by pretreatment with 2-deoxyglucose and sodium azide, which results in ATP depletion, but not by pretreatment with NaF and AlCl3, which activates heterotrimeric G proteins. Moreover, we found that palmitoylation of the UL51 protein through the N-terminal cysteine at position 9 was necessary for its Golgi localization. Protease digestion analysis suggested that the UL51 protein localized on the cytoplasmic face of the membrane in UL51-transfected cells, while in infected cells it localized mainly to the inside of cytoplasmic vesicles and/or the viral envelope. Transmission immunoelectron microscopy revealed an association of UL51 protein-specific labeling with cytoplasmic virions and also with some membranous structure. We infer from these observations that internalization of UL51 protein into the cytoplasmic vesicle and/or virion may occur in association with viral envelonment in HSV-infected cells.

Original languageEnglish
Pages (from-to)3204-3216
Number of pages13
JournalJournal of Virology
Volume77
Issue number5
DOIs
Publication statusPublished - 01-03-2003
Externally publishedYes

Fingerprint

Lipoylation
Human herpesvirus 1
Human Herpesvirus 1
Golgi Apparatus
Golgi apparatus
Proteins
proteins
Virion
Cytoplasmic Vesicles
virion
cells
palmitoylation
pretreatment
Brefeldin A
Heterotrimeric GTP-Binding Proteins
2-deoxyglucose
Sodium Azide
brefeldin A
herpes simplex
Immunoelectron Microscopy

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Nozawa, Naoki ; Daikoku, Tohru ; Koshizuka, Tetsuo ; Yamauchi, Yohei ; Yoshikawa, Tetsushi ; Nishiyama, Yukihiro. / Subcellular localization of herpes simplex virus type 1 UL51 protein and role of palmitoylation in golgi apparatus targeting. In: Journal of Virology. 2003 ; Vol. 77, No. 5. pp. 3204-3216.
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abstract = "The herpes simplex virus type I (HSV-1) UL51 gene products are virion-associated phosphoproteins with apparent molecular masses of 27, 29, and 30 kDa in HSV-1-infected cells. In this study, we have investigated the intracellular localization and distribution of UL51 protein both in infected cells and in transfected cells expressing only UL51. We found that this protein colocalized closely with Golgi marker proteins such as the Golgi-58K protein and GM130 in transfected cells expressing only UL51. However, in infected cells, the UL51 protein localized to the juxtanuclear region but only partially colocalized with the Golgi maker proteins. Mutant protein analysis revealed that the N-terminal 15 amino acid residues of the UL51 protein sufficed for this Golgi localization property. The UL51 protein redistributed on addition of brefeldin A. This was prevented by pretreatment with 2-deoxyglucose and sodium azide, which results in ATP depletion, but not by pretreatment with NaF and AlCl3, which activates heterotrimeric G proteins. Moreover, we found that palmitoylation of the UL51 protein through the N-terminal cysteine at position 9 was necessary for its Golgi localization. Protease digestion analysis suggested that the UL51 protein localized on the cytoplasmic face of the membrane in UL51-transfected cells, while in infected cells it localized mainly to the inside of cytoplasmic vesicles and/or the viral envelope. Transmission immunoelectron microscopy revealed an association of UL51 protein-specific labeling with cytoplasmic virions and also with some membranous structure. We infer from these observations that internalization of UL51 protein into the cytoplasmic vesicle and/or virion may occur in association with viral envelonment in HSV-infected cells.",
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Subcellular localization of herpes simplex virus type 1 UL51 protein and role of palmitoylation in golgi apparatus targeting. / Nozawa, Naoki; Daikoku, Tohru; Koshizuka, Tetsuo; Yamauchi, Yohei; Yoshikawa, Tetsushi; Nishiyama, Yukihiro.

In: Journal of Virology, Vol. 77, No. 5, 01.03.2003, p. 3204-3216.

Research output: Contribution to journalArticle

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AU - Nozawa, Naoki

AU - Daikoku, Tohru

AU - Koshizuka, Tetsuo

AU - Yamauchi, Yohei

AU - Yoshikawa, Tetsushi

AU - Nishiyama, Yukihiro

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