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REPLICATION

The Zinc Finger Antiviral Protein Directly Binds to Specific Viral mRNAs through the CCCH Zinc Finger Motifs

Xuemin Guo, John-William N. Carroll, Margaret R. MacDonald, Stephen P. Goff, Guangxia Gao
Xuemin Guo
1Institute of Microbiology
2Graduate School, Chinese Academy of Sciences, Beijing, China
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John-William N. Carroll
3Laboratory of Virology and Infectious Diseases, The Rockefeller University
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Margaret R. MacDonald
3Laboratory of Virology and Infectious Diseases, The Rockefeller University
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Stephen P. Goff
4Department of Biochemistry and Molecular Biophysics, Howard Hughes Medical Institute, Columbia University, New York, New York
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Guangxia Gao
1Institute of Microbiology
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  • For correspondence: gaogx@sun.im.ac.cn
DOI: 10.1128/JVI.78.23.12781-12787.2004
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ABSTRACT

The zinc finger antiviral protein (ZAP) is a recently isolated host antiviral factor. It specifically inhibits the replication of Moloney murine leukemia virus (MLV) and Sindbis virus (SIN) by preventing the accumulation of viral RNA in the cytoplasm. For this report, we mapped the viral sequences that are sensitive to ZAP inhibition. The viral sequences were cloned into a luciferase reporter and analyzed for the ability to mediate ZAP-dependent destabilization of the reporter. The sensitive sequence in MLV was mapped to the 3′ long terminal repeat; the sensitive sequences in SIN were mapped to multiple fragments. The fragment of SIN that displayed the highest destabilizing activity was further analyzed by deletion mutagenesis for the minimal sequence that retained the activity. This led to the identification of a fragment of 653 nucleotides. Any further deletion of this fragment resulted in significantly lower activity. We provide evidence that ZAP directly binds to the active but not the inactive fragments. The CCCH zinc finger motifs of ZAP play important roles in RNA binding and antiviral activity. Disruption of the second and fourth zinc fingers abolished ZAP's activity, whereas disruption of the first and third fingers just slightly lowered its activity.

  • Copyright © 2004 American Society for Microbiology
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The Zinc Finger Antiviral Protein Directly Binds to Specific Viral mRNAs through the CCCH Zinc Finger Motifs
Xuemin Guo, John-William N. Carroll, Margaret R. MacDonald, Stephen P. Goff, Guangxia Gao
Journal of Virology Nov 2004, 78 (23) 12781-12787; DOI: 10.1128/JVI.78.23.12781-12787.2004

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The Zinc Finger Antiviral Protein Directly Binds to Specific Viral mRNAs through the CCCH Zinc Finger Motifs
Xuemin Guo, John-William N. Carroll, Margaret R. MacDonald, Stephen P. Goff, Guangxia Gao
Journal of Virology Nov 2004, 78 (23) 12781-12787; DOI: 10.1128/JVI.78.23.12781-12787.2004
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KEYWORDS

Carrier Proteins
RNA, Messenger
RNA, Viral
Zinc Fingers

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