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Journal of Virology, March 2008, p. 2286-2294, Vol. 82, No. 5
0022-538X/08/$08.00+0     doi:10.1128/JVI.01761-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

New mRNAs Are Preferentially Translated during Vesicular Stomatitis Virus Infection

Zackary W. Whitlow, John H. Connor, and Douglas S. Lyles^*

Department of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27157

Received 11 August 2007/ Accepted 4 December 2007

During vesicular stomatitis virus (VSV) infection, host protein synthesis is inhibited, while synthesis of viral proteins increases. VSV infection causes inhibition of host transcription and RNA transport. Therefore, most host mRNAs in the cytoplasm of infected cells were synthesized before infection. However, viral mRNAs are synthesized throughout infection and are newer than preexisting host mRNAs. To determine if the timing of appearance of mRNAs in the cytoplasm affected their translation during VSV infection, we transfected reporter mRNAs into cells at various times relative to the time of infection and measured their rate of translation in mock- and VSV-infected cells. We found that translation of mRNAs transfected during infection was not inhibited but that translation of mRNAs transfected prior to infection was inhibited during VSV infection. Based on these data, we conclude that the timing of viral mRNA appearance in the cytoplasm is responsible, at least in part, for the preferential translation of VSV mRNAs. A time course measuring translation efficiencies of viral and host mRNAs showed that the translation efficiencies of viral mRNAs increased between 4 and 8 h postinfection, while translation efficiencies of host mRNAs decreased. The increased translation efficiency of viral mRNAs occurred in cells infected with an M protein mutant virus that is defective in host shutoff, demonstrating that the enhanced translation of viral mRNA is genetically separable from inhibition of translation of host mRNA.

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* Corresponding author. Mailing address: Department of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, NC 27157. Phone: (336) 716-4237. Fax: (336) 716-7671. E-mail: dlyles{at}wfubmc.edu

Published ahead of print on 19 December 2007.

Present address: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892.

Present address: Department of Microbiology, Boston University School of Medicine, Boston, MA 02118.

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Journal of Virology, March 2008, p. 2286-2294, Vol. 82, No. 5
0022-538X/08/$08.00+0     doi:10.1128/JVI.01761-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

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