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Zinc-finger antiviral protein inhibits XMRV infection.

Wang X, Tu F, Zhu Y, Gao G - PLoS ONE (2012)

Bottom Line: ZAP inhibits XMRV replication by preventing the accumulation of viral mRNA in the cytoplasm.Documentation of ZAP inhibiting XMRV helps to broaden the spectrum of ZAP's antiviral activity.Comparison of the target sequences of ZAP in XMRV and MoMLV helps to better understand the features of ZAP-responsive elements.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Infection and Immunity, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.

ABSTRACT

Background: The zinc-finger antiviral protein (ZAP) is a host factor that specifically inhibits the replication of certain viruses, including Moloney murine leukemia virus (MoMLV), HIV-1, and certain alphaviruses and filoviruses. ZAP binds to specific viral mRNAs and recruits cellular mRNA degradation machinery to degrade the target RNA. The common features of ZAP-responsive RNA sequences remain elusive and thus whether a virus is susceptible to ZAP can only be determined experimentally. Xenotropic murine leukemia virus-related virus (XMRV) is a recently identified γ-retrovirus that was originally thought to be involved in prostate cancer and chronic fatigue syndrome but recently proved to be a laboratory artefact. Nonetheless, XMRV as a new retrovirus has been extensively studied. Since XMRV and MoMLV share only 67.9% sequence identity in the 3'UTRs, which is the target sequence of ZAP in MoMLV, whether XMRV is susceptible to ZAP remains to be determined.

Findings: We constructed an XMRV-luc vector, in which the coding sequences of Gag-Pol and part of Env were replaced with luciferase-coding sequence. Overexpression of ZAP potently inhibited the expression of XMRV-luc in a ZAP expression-level-dependent manner, while downregulation of endogenous ZAP rendered cells more sensitive to infection. Furthermore, ZAP inhibited the spreading of replication-competent XMRV. Consistent with the previously reported mechanisms by which ZAP inhibits viral infection, ZAP significantly inhibited the accumulation of XMRV-luc mRNA in the cytoplasm. The ZAP-responsive element in XMRV mRNA was mapped to the 3'UTR.

Conclusions: ZAP inhibits XMRV replication by preventing the accumulation of viral mRNA in the cytoplasm. Documentation of ZAP inhibiting XMRV helps to broaden the spectrum of ZAP's antiviral activity. Comparison of the target sequences of ZAP in XMRV and MoMLV helps to better understand the features of ZAP-responsive elements.

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Mapping of ZRE in the 3′ LTR of XMRV. (A) Schematic structures of the truncation constructs of 3′UTR. (B) Analysis of the sensitivity of the 3′UTR truncation mutants to ZAP. Fold inhibition was measured as described in the legend to Figure 5B. Data presented are means ± SD of three independent experiments.
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pone-0039159-g006: Mapping of ZRE in the 3′ LTR of XMRV. (A) Schematic structures of the truncation constructs of 3′UTR. (B) Analysis of the sensitivity of the 3′UTR truncation mutants to ZAP. Fold inhibition was measured as described in the legend to Figure 5B. Data presented are means ± SD of three independent experiments.

Mentions: The ZRE in MoMLV was also mapped to the 3′UTR [5]. Sequence analysis reveals that the 3′UTRs of XMRV and MoMLV share 67.9% identity (Fig. S2). To map the minimal sequence required for response to ZAP, the 3′UTR of XMRV was truncated and the truncation mutants were analyzed for their sensitivity to ZAP (Fig. 6A). The above identified XMRV 3′UTR covers a short fragment of env, U3 and the R region. Deletion of the env sequence and R region did not significantly affect the sensitivity to ZAP (Fig. 6B). However, further deletion resulted in a significant drop in the sensitivity (Fig. 6B), suggesting that the fragment covering the U3 region is the ZRE in XMRV.


Zinc-finger antiviral protein inhibits XMRV infection.

Wang X, Tu F, Zhu Y, Gao G - PLoS ONE (2012)

Mapping of ZRE in the 3′ LTR of XMRV. (A) Schematic structures of the truncation constructs of 3′UTR. (B) Analysis of the sensitivity of the 3′UTR truncation mutants to ZAP. Fold inhibition was measured as described in the legend to Figure 5B. Data presented are means ± SD of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3376128&req=5

pone-0039159-g006: Mapping of ZRE in the 3′ LTR of XMRV. (A) Schematic structures of the truncation constructs of 3′UTR. (B) Analysis of the sensitivity of the 3′UTR truncation mutants to ZAP. Fold inhibition was measured as described in the legend to Figure 5B. Data presented are means ± SD of three independent experiments.
Mentions: The ZRE in MoMLV was also mapped to the 3′UTR [5]. Sequence analysis reveals that the 3′UTRs of XMRV and MoMLV share 67.9% identity (Fig. S2). To map the minimal sequence required for response to ZAP, the 3′UTR of XMRV was truncated and the truncation mutants were analyzed for their sensitivity to ZAP (Fig. 6A). The above identified XMRV 3′UTR covers a short fragment of env, U3 and the R region. Deletion of the env sequence and R region did not significantly affect the sensitivity to ZAP (Fig. 6B). However, further deletion resulted in a significant drop in the sensitivity (Fig. 6B), suggesting that the fragment covering the U3 region is the ZRE in XMRV.

Bottom Line: ZAP inhibits XMRV replication by preventing the accumulation of viral mRNA in the cytoplasm.Documentation of ZAP inhibiting XMRV helps to broaden the spectrum of ZAP's antiviral activity.Comparison of the target sequences of ZAP in XMRV and MoMLV helps to better understand the features of ZAP-responsive elements.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Infection and Immunity, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.

ABSTRACT

Background: The zinc-finger antiviral protein (ZAP) is a host factor that specifically inhibits the replication of certain viruses, including Moloney murine leukemia virus (MoMLV), HIV-1, and certain alphaviruses and filoviruses. ZAP binds to specific viral mRNAs and recruits cellular mRNA degradation machinery to degrade the target RNA. The common features of ZAP-responsive RNA sequences remain elusive and thus whether a virus is susceptible to ZAP can only be determined experimentally. Xenotropic murine leukemia virus-related virus (XMRV) is a recently identified γ-retrovirus that was originally thought to be involved in prostate cancer and chronic fatigue syndrome but recently proved to be a laboratory artefact. Nonetheless, XMRV as a new retrovirus has been extensively studied. Since XMRV and MoMLV share only 67.9% sequence identity in the 3'UTRs, which is the target sequence of ZAP in MoMLV, whether XMRV is susceptible to ZAP remains to be determined.

Findings: We constructed an XMRV-luc vector, in which the coding sequences of Gag-Pol and part of Env were replaced with luciferase-coding sequence. Overexpression of ZAP potently inhibited the expression of XMRV-luc in a ZAP expression-level-dependent manner, while downregulation of endogenous ZAP rendered cells more sensitive to infection. Furthermore, ZAP inhibited the spreading of replication-competent XMRV. Consistent with the previously reported mechanisms by which ZAP inhibits viral infection, ZAP significantly inhibited the accumulation of XMRV-luc mRNA in the cytoplasm. The ZAP-responsive element in XMRV mRNA was mapped to the 3'UTR.

Conclusions: ZAP inhibits XMRV replication by preventing the accumulation of viral mRNA in the cytoplasm. Documentation of ZAP inhibiting XMRV helps to broaden the spectrum of ZAP's antiviral activity. Comparison of the target sequences of ZAP in XMRV and MoMLV helps to better understand the features of ZAP-responsive elements.

Show MeSH
Related in: MedlinePlus