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Journal of Virology, January 2009, p. 770-780, Vol. 83, No. 2
0022-538X/09/$08.00+0     doi:10.1128/JVI.01279-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

hnRNPs Relocalize to the Cytoplasm following Infection with Vesicular Stomatitis Virus{triangledown}

Elizabeth L. Pettit Kneller,1 John H. Connor,2 and Douglas S. Lyles1*

Department of Biochemistry, Wake Forest University School of Medicine, Medical Center Boulevard, Winston-Salem, North Carolina 27157,1 Department of Microbiology, Boston University School of Medicine, 75 East Concord St., Boston, Massachusetts 021182

Received 19 June 2008/ Accepted 28 October 2008

Vesicular stomatitis virus (VSV) matrix protein inhibits nuclear-cytoplasmic mRNA transport. The goal of this work is to determine whether VSV inhibits the nuclear-cytoplasmic transport of heterogeneous ribonucleoproteins (hnRNPs), which are thought to serve as mRNA export factors. Confocal microscopy experiments showed that hnRNPA1, hnRNPK, and hnRNPC1/C2, but not hnRNPB1 or lamin A/C, are relocalized to the cytoplasm during VSV infection. We determined whether protein import is inhibited by VSV by transfecting cells with a plasmid encoding enhanced green fluorescent protein (EGFP) tagged with either the M9 nuclear localization sequence (NLS) or the classical NLS. These experiments revealed that both the M9 NLS and the classical NLS are functional during VSV infection. These data suggest that the inhibition of protein import is not responsible for hnRNP relocalization during VSV infection but that hnRNP export is enhanced. We found that hnRNPA1 relocalization was significantly reduced following the silencing of the mRNA export factor Rae1, indicating that Rae1 is necessary for hnRNP export. In order to determine the role of hnRNPA1 in VSV infection, we silenced hnRNPA1 in HeLa cells and assayed three aspects of the viral life cycle: host protein synthesis shutoff concurrent with the onset of viral protein synthesis, replication by plaque assay, and cell killing. We observed that host shutoff and replication are unaffected by the reduction in hnRNPA1 but that the rate of VSV-induced apoptosis is slower in cells that have reduced hnRNPA1. These data suggest that VSV promotes hnRNPA1 relocalization in a Rae1-dependent manner for apoptotic signaling.

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

{triangledown} Published ahead of print on 12 November 2008.

Journal of Virology, January 2009, p. 770-780, Vol. 83, No. 2
0022-538X/09/$08.00+0     doi:10.1128/JVI.01279-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



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  • Lin, J.-Y., Shih, S.-R., Pan, M., Li, C., Lue, C.-F., Stollar, V., Li, M.-L. (2009). hnRNP A1 Interacts with the 5' Untranslated Regions of Enterovirus 71 and Sindbis Virus RNA and Is Required for Viral Replication. J. Virol. 83: 6106-6114 [Abstract] [Full Text]  


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