Effect of USP9X on HSV-1 replication. (A) HeLa cells ex

Ubiquitin-specific protease 9X in host cells interacts with herpes simplex virus 1 ICP0. Sato Y, Kato A, Arii J, Koyanagi N, Kozuka-Hata H, Oyama M, Kawaguchi Y - J. Vet. Med. Sci. (2015) Bottom Line: The interaction between ICP0 and USP9X was confirmed by co-immunoprecipitation.Notably, USP9X depletion increased the ICP0 abundance and promoted viral replication.These results suggest that USP9X-dependent regulation of ICP0 expression is part of a complex feedback mechanism that facilitates optimal HSV-1 replication. View Article: PubMed Central - PubMed Affiliation: Division of Molecular Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan. ABSTRACT Herpes simplex virus 1 (HSV-1) expresses infected cell protein 0 (ICP0), a multi-functional protein with E3 ubiquitin ligase activity and a critical regulator of the viral life cycle. To obtain novel insights into the molecular mechanism by which ICP0 regulates HSV-1 replication, we analyzed HEp-2 cells infected with HSV-1 by tandem affinity purification and mass spectrometry-based proteomics. This screen identified 50 host-cell proteins that potentially interact with ICP0, including ubiquitin-specific protease 9X (USP9X). The interaction between ICP0 and USP9X was confirmed by co-immunoprecipitation. Notably, USP9X depletion increased the ICP0 abundance and promoted viral replication. These results suggest that USP9X-dependent regulation of ICP0 expression is part of a complex feedback mechanism that facilitates optimal HSV-1 replication. No MeSH data available. Related in: MedlinePlus	© Copyright Policy - open-access Related In: Results - Collection License Show All Figures getmorefigures.php?uid=PMC4829507&req=5 fig_005: Effect of USP9X on HSV-1 replication. (A) HeLa cells expressing silencing shRNAagainst LacZ and USP9X were analyzed by immunoblotting with antibodies against USP9Xand β-actin. (B) These cells were infected with wild-type HSV-1 (F) at MOI 0.01. Totalvirus preparations from the media and infected cells were obtained 48 hr postinfectionand assayed on U-2 OS cells. Data are mean ± standard error from three independentexperiments. P<0.05 by two-tailed Student’st-test. Mentions:* Effect of USP9X on HSV-1 replication: To investigate the role (s) of USP9Xin HSV-1 replication, we generated HeLa cell lines stably expressing silencing shRNA againstUSP9X, as well as a control cell line expressing shRNA against LacZ. Expression of USP9X wasconsiderably diminished in cells expressing USP9X shRNA than in cells with LacZ shRNA (Fig. 5AFig. 5.

Ubiquitin-specific protease 9X in host cells interacts with herpes simplex virus 1 ICP0.

Sato Y, Kato A, Arii J, Koyanagi N, Kozuka-Hata H, Oyama M, Kawaguchi Y - J. Vet. Med. Sci. (2015)

Bottom Line: The interaction between ICP0 and USP9X was confirmed by co-immunoprecipitation.Notably, USP9X depletion increased the ICP0 abundance and promoted viral replication.These results suggest that USP9X-dependent regulation of ICP0 expression is part of a complex feedback mechanism that facilitates optimal HSV-1 replication.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Virology, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan.

ABSTRACT

Herpes simplex virus 1 (HSV-1) expresses infected cell protein 0 (ICP0), a multi-functional protein with E3 ubiquitin ligase activity and a critical regulator of the viral life cycle. To obtain novel insights into the molecular mechanism by which ICP0 regulates HSV-1 replication, we analyzed HEp-2 cells infected with HSV-1 by tandem affinity purification and mass spectrometry-based proteomics. This screen identified 50 host-cell proteins that potentially interact with ICP0, including ubiquitin-specific protease 9X (USP9X). The interaction between ICP0 and USP9X was confirmed by co-immunoprecipitation. Notably, USP9X depletion increased the ICP0 abundance and promoted viral replication. These results suggest that USP9X-dependent regulation of ICP0 expression is part of a complex feedback mechanism that facilitates optimal HSV-1 replication.

No MeSH data available.

Related in: MedlinePlus

Effect of USP9X on HSV-1 replication. (A) HeLa cells expressing silencing shRNAagainst LacZ and USP9X were analyzed by immunoblotting with antibodies against USP9Xand β-actin. (B) These cells were infected with wild-type HSV-1 (F) at MOI 0.01. Totalvirus preparations from the media and infected cells were obtained 48 hr postinfectionand assayed on U-2 OS cells. Data are mean ± standard error from three independentexperiments. *P<0.05 by two-tailed Student’st-test.

Related In: Results - Collection

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fig_005: Effect of USP9X on HSV-1 replication. (A) HeLa cells expressing silencing shRNAagainst LacZ and USP9X were analyzed by immunoblotting with antibodies against USP9Xand β-actin. (B) These cells were infected with wild-type HSV-1 (F) at MOI 0.01. Totalvirus preparations from the media and infected cells were obtained 48 hr postinfectionand assayed on U-2 OS cells. Data are mean ± standard error from three independentexperiments. *P<0.05 by two-tailed Student’st-test.

Mentions: Effect of USP9X on HSV-1 replication: To investigate the role (s) of USP9Xin HSV-1 replication, we generated HeLa cell lines stably expressing silencing shRNA againstUSP9X, as well as a control cell line expressing shRNA against LacZ. Expression of USP9X wasconsiderably diminished in cells expressing USP9X shRNA than in cells with LacZ shRNA (Fig. 5AFig. 5.