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Journal of Virology, May 1999, p. 4524-4529, Vol. 73, No. 5
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification and Initial Characterization of the Murine Gammaherpesvirus 68 Gene M3, Encoding an Abundantly Secreted Protein

Victor van Berkel, Karen Preiter, Herbert W. Virgin IV,* and Samuel H. Speck*

Center for Immunology, and Departments of Pathology and Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri

Received 4 December 1998/Accepted 11 February 1999

Several viruses, including members of the gammaherpesvirus family, encode proteins that are secreted into the extracellular environment. We have identified an abundant 44-kDa secreted protein that is present in the supernatant of fibroblasts infected with murine gammaherpesvirus 68 (gamma HV68; also referred to as MHV-68) but not in that of uninfected fibroblasts. Sequence analysis of the amino terminus and of internal peptides revealed that this protein is encoded by the gamma HV68 M3 open reading frame (ORF). The amino-terminal sequence of the secreted protein starts at residue 25 of the M3 ORF, consistent with the first 24 residues functioning as a signal peptide. Northern blot analysis revealed a single abundant ~1.4-kb early-late lytic transcript encoded by the M3 ORF. Analysis of a partial cDNA clone and subsequent analyses of products of rapid amplification of cDNA ends coupled with S1 nuclease protection assays demonstrate that the M3 protein is encoded by an unspliced, polyadenylated mRNA initiating at bp 7294 and terminating at bp 6007 of the gamma HV68 genome. The 3' end of the M3 transcript maps 9 bp downstream of a consensus polyadenylation signal. Thus, the predicted M3 ORF is a functional gene that encodes an abundant secreted protein which is a candidate for interacting with host cellular receptors or cytokines.

* Corresponding author. Mailing address: Department of Pathology, Box 8118, Washington University in St. Louis, 660 South Euclid Ave., St. Louis, MO 63110-1093. Phone: (314) 362-9223 (for H.W.V.) and (314) 362-0367 (for S.H.S.). Fax: (314) 362-4096. E-mail: virgin{at}immunology.wustl.edu (for H.W.V.) and speck{at}pathbox.wustl.edu (for S.H.S.).

Journal of Virology, May 1999, p. 4524-4529, Vol. 73, No. 5
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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