The UL3 open reading frame of herpes simplex virus type 1 codes for a phosphoprotein

Homayon Ghiasi, Guey Chuen Perng, Steve Cai, Anthony B. Nesburn, Steven L. Wechsler

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Based on sequence analysis, the protein encoded by the UL3 open reading frame (ORF) of herpes simplex virus type 1 (HSV-1) was predicted to contain an N glucosylation site and to be a glycoprotein. To determine if this prediction was correct, we cloned and expressed the DNA encoding the complete sequence of the UL3 ORF in a baculovirus expression system. Western blotting was done using polyclonal antibody raised against synthetic UL3 peptides. Two major baculovirus-UL3 expressed protein bands with apparent molecular weights of 30 kDa and 31 kDa, and two minor protein bands with apparent molecular weights of 29 kDa and 33 kDa were detected. None of the expressed UL3 protein species were susceptible to tunicamycin treatment, suggesting that they were not N-linked glycosy]ated. Cell fractionation studies indicated that the UL3 protein was localized in the cytoplasmic and nuclear portion of the cells, rather than the cell membrane, again suggesting a lack of glycosylation. In contrast, the baculovirus expressed UL3 protein was phosphorylated as judged by 32P(i)-labeling. Immunoprecipitation followed by SDS-PAGE demonstrated a single 32P(i)-labeled UL3 related band with an apparent molecular weight of 33 kDa, indicating that the UL3 protein was a phosphoprotein. Antibodies produced in mice vaccinated with baculovirus-U13 protein reacted with two UL3 related HSV-1 bands on Western blots. These protein bands had apparent molecular weights of 27 and 33 kDa and presumably represent the unphosphorylated and phosphorylated forms of UL3.

Original languageEnglish
Pages (from-to)137-142
Number of pages6
JournalVirus Research
Issue number2
Publication statusPublished - 1996 Oct

All Science Journal Classification (ASJC) codes

  • Cancer Research
  • Virology
  • Infectious Diseases


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