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Herpes simplex virus 1 UL41 protein abrogates the antiviral activity of hZAP by degrading its mRNA.

Su C, Zhang J, Zheng C - Virol. J. (2015)

Bottom Line: However, previous study demonstrated that ectopic expression of rat ZAP did not suppress the replication of herpes simplex virus type 1 (HSV-1), an archetypal member of the alphaherpesvirus subfamily, and the molecular mechanism underneath is still illusive.Moreover, ectopic expression of hZAP inhibited the replication of R2621 but not WT HSV-1.HSV-1 UL41 was shown for the first time to evade the antiviral function of hZAP via its RNase activity.

View Article: PubMed Central - PubMed

Affiliation: Institutes of Biology and Medical Sciences, Soochow University, Suzhou, 215123, China. such1986@aliyun.com.

ABSTRACT

Background: The zinc finger antiviral protein (ZAP) is a host restriction factor that inhibits the replication of various viruses by degradation of certain viral mRNA. However, previous study demonstrated that ectopic expression of rat ZAP did not suppress the replication of herpes simplex virus type 1 (HSV-1), an archetypal member of the alphaherpesvirus subfamily, and the molecular mechanism underneath is still illusive.

Results: Human ZAP (hZAP) does not suppress the replication of herpes simplex virus 1, and HSV-1 UL41 protein was identified as an antagonist of hZAP by degrading its mRNA. Infection of wild-type (WT), but not UL41- mutant (R2621) virus, diminished the accumulation of hZAP to abrogate its antiviral activity. Moreover, ectopic expression of hZAP inhibited the replication of R2621 but not WT HSV-1.

Conclusion: HSV-1 UL41 was shown for the first time to evade the antiviral function of hZAP via its RNase activity.

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Related in: MedlinePlus

HSV-1 UL41 protein abrogated the antiviral activity of hZAP. a and b 293Trex-hZAPL cells and 293Trex-hZAPS cells were infected with HSV-1-BAC-Luc at an MOI of 1. Cells were treated or mock treated with tetracycline (100 ng/mL or 1000 ng/mL) 2 h post-infection to induce hZAP expression. The cells were lysed, and luciferase activities were measured at 36 h after infection. Fold inhibition was calculated as the ratio of the luciferase activity in mock-treated cells to that in tetracycline-treated cells. c 293Trex-hZAPS cells were transfected with 500 ng of NL4-3-Luc reporter plasmid, together with Renilla luciferase plasmid pRL-TK (50 ng) and pEYFP-N1 control vector or plasmids encoding the indicated viral proteins (1000 ng). At 6 h post-transfection, cells were treated with or without tetracycline (1000 ng/mL) to induce hZAPS expression and were incubated for additional 36 h, followed by cell lysis. The luciferase activity was determined by a dual-luciferase assay. Data shown are means ± SD from three separate experiments. (*P < 0.05)
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Fig1: HSV-1 UL41 protein abrogated the antiviral activity of hZAP. a and b 293Trex-hZAPL cells and 293Trex-hZAPS cells were infected with HSV-1-BAC-Luc at an MOI of 1. Cells were treated or mock treated with tetracycline (100 ng/mL or 1000 ng/mL) 2 h post-infection to induce hZAP expression. The cells were lysed, and luciferase activities were measured at 36 h after infection. Fold inhibition was calculated as the ratio of the luciferase activity in mock-treated cells to that in tetracycline-treated cells. c 293Trex-hZAPS cells were transfected with 500 ng of NL4-3-Luc reporter plasmid, together with Renilla luciferase plasmid pRL-TK (50 ng) and pEYFP-N1 control vector or plasmids encoding the indicated viral proteins (1000 ng). At 6 h post-transfection, cells were treated with or without tetracycline (1000 ng/mL) to induce hZAPS expression and were incubated for additional 36 h, followed by cell lysis. The luciferase activity was determined by a dual-luciferase assay. Data shown are means ± SD from three separate experiments. (*P < 0.05)

Mentions: Previous study showed that ectopic expression of rat ZAP did not affect HSV-1 infection [16]. To investigate whether hZAP could inhibit HSV-1 infection, 293TRex-hZAPL/ZAPS cells, which expressed ZAPL/ZAPS in a tetracycline (Tet)-inducible manner, were infected with HSV-1-BAC-Luc at an MOI of 0.1, 1 or 10 and then mock-treated or treated with 100 ng/mL or 1000 ng/mL Tet [12, 17]. Luciferase activity was measured to determine the replication of viruses. As a result, different MOI of HSV-1 replicated similarly in mock and hZAPL- or hZAPS-expressing cells (Fig. 1a, b and data not shown). Viral plaque assay was also conducted and similar result was obtained (data not shown). These results demonstrate that hZAP does not inhibit HSV-1 infection.Fig. 1


Herpes simplex virus 1 UL41 protein abrogates the antiviral activity of hZAP by degrading its mRNA.

Su C, Zhang J, Zheng C - Virol. J. (2015)

HSV-1 UL41 protein abrogated the antiviral activity of hZAP. a and b 293Trex-hZAPL cells and 293Trex-hZAPS cells were infected with HSV-1-BAC-Luc at an MOI of 1. Cells were treated or mock treated with tetracycline (100 ng/mL or 1000 ng/mL) 2 h post-infection to induce hZAP expression. The cells were lysed, and luciferase activities were measured at 36 h after infection. Fold inhibition was calculated as the ratio of the luciferase activity in mock-treated cells to that in tetracycline-treated cells. c 293Trex-hZAPS cells were transfected with 500 ng of NL4-3-Luc reporter plasmid, together with Renilla luciferase plasmid pRL-TK (50 ng) and pEYFP-N1 control vector or plasmids encoding the indicated viral proteins (1000 ng). At 6 h post-transfection, cells were treated with or without tetracycline (1000 ng/mL) to induce hZAPS expression and were incubated for additional 36 h, followed by cell lysis. The luciferase activity was determined by a dual-luciferase assay. Data shown are means ± SD from three separate experiments. (*P < 0.05)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4666169&req=5

Fig1: HSV-1 UL41 protein abrogated the antiviral activity of hZAP. a and b 293Trex-hZAPL cells and 293Trex-hZAPS cells were infected with HSV-1-BAC-Luc at an MOI of 1. Cells were treated or mock treated with tetracycline (100 ng/mL or 1000 ng/mL) 2 h post-infection to induce hZAP expression. The cells were lysed, and luciferase activities were measured at 36 h after infection. Fold inhibition was calculated as the ratio of the luciferase activity in mock-treated cells to that in tetracycline-treated cells. c 293Trex-hZAPS cells were transfected with 500 ng of NL4-3-Luc reporter plasmid, together with Renilla luciferase plasmid pRL-TK (50 ng) and pEYFP-N1 control vector or plasmids encoding the indicated viral proteins (1000 ng). At 6 h post-transfection, cells were treated with or without tetracycline (1000 ng/mL) to induce hZAPS expression and were incubated for additional 36 h, followed by cell lysis. The luciferase activity was determined by a dual-luciferase assay. Data shown are means ± SD from three separate experiments. (*P < 0.05)
Mentions: Previous study showed that ectopic expression of rat ZAP did not affect HSV-1 infection [16]. To investigate whether hZAP could inhibit HSV-1 infection, 293TRex-hZAPL/ZAPS cells, which expressed ZAPL/ZAPS in a tetracycline (Tet)-inducible manner, were infected with HSV-1-BAC-Luc at an MOI of 0.1, 1 or 10 and then mock-treated or treated with 100 ng/mL or 1000 ng/mL Tet [12, 17]. Luciferase activity was measured to determine the replication of viruses. As a result, different MOI of HSV-1 replicated similarly in mock and hZAPL- or hZAPS-expressing cells (Fig. 1a, b and data not shown). Viral plaque assay was also conducted and similar result was obtained (data not shown). These results demonstrate that hZAP does not inhibit HSV-1 infection.Fig. 1

Bottom Line: However, previous study demonstrated that ectopic expression of rat ZAP did not suppress the replication of herpes simplex virus type 1 (HSV-1), an archetypal member of the alphaherpesvirus subfamily, and the molecular mechanism underneath is still illusive.Moreover, ectopic expression of hZAP inhibited the replication of R2621 but not WT HSV-1.HSV-1 UL41 was shown for the first time to evade the antiviral function of hZAP via its RNase activity.

View Article: PubMed Central - PubMed

Affiliation: Institutes of Biology and Medical Sciences, Soochow University, Suzhou, 215123, China. such1986@aliyun.com.

ABSTRACT

Background: The zinc finger antiviral protein (ZAP) is a host restriction factor that inhibits the replication of various viruses by degradation of certain viral mRNA. However, previous study demonstrated that ectopic expression of rat ZAP did not suppress the replication of herpes simplex virus type 1 (HSV-1), an archetypal member of the alphaherpesvirus subfamily, and the molecular mechanism underneath is still illusive.

Results: Human ZAP (hZAP) does not suppress the replication of herpes simplex virus 1, and HSV-1 UL41 protein was identified as an antagonist of hZAP by degrading its mRNA. Infection of wild-type (WT), but not UL41- mutant (R2621) virus, diminished the accumulation of hZAP to abrogate its antiviral activity. Moreover, ectopic expression of hZAP inhibited the replication of R2621 but not WT HSV-1.

Conclusion: HSV-1 UL41 was shown for the first time to evade the antiviral function of hZAP via its RNase activity.

Show MeSH
Related in: MedlinePlus