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Intracellular Localization of Polypeptides Encoded in Mouse Hepatitis Virus Open Reading Frame 1A

  • Weizhen Bi
  • Pedro J. Bonilla
  • Kathryn V. Holmes
  • Susan R. Weiss
  • Julian L. Leibowitz
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 380)

Abstract

We have investigated the intracellular localization of several of the proteolytic cleavage products derived from the 5′ portion of mouse hepatitis virus (MHV) gene 1. Antisera UP1 recognizes the N-terminal ORF la cleavage product p28. Immunofluorescent staining of cells with this antisera resulted in a diffuse punctate pattern of cytoplasmic staining, indicating that this protein is widely distributed in the cytoplasm. Immunofluorescent staining of infected cells with antisera which recognize polypeptides p240 and p290 stained discrete vesicular perinuclear structures suggesting that these proteins localized to the Golgi. This was confirmed by double immunofluorescent staining of BHK cells expressing the MHV receptor (BHK-R) with a Golgi specific antibody in addition to our anti-MHV ORF 1a antibodies. Antisera UP 102 recognizes p28 and the immediately downstream p65 gene product. Double immunofluorescent staining of MHV infected BHK-R cells with UP 102 labeled discrete vesicular structures overlapping the Golgi complex. In addition there was punctate staining more widely distributed in the cytoplasm. The simplest explanation for this pattern is that p65 is also localized to the Golgi region of the cell, whereas p28 is more widespread. Plasmids containing the first 4.7 and 6.75 kb of ORF la have been expressed using the coupled vaccinia virus — T7 polymerase system. Images obtained by immunofluorescent staining of transfectants with our anti-ORF1a antisera are similar to those obtained during infection with A59. These studies indicate that the signals which direct p290 to the Golgi are likely contained between the C-terminus of p28 and ORF la residue 1494.

Keywords

Golgi Complex Double Immunofluorescent Staining Mouse Hepatitis Virus Immunofluorescent Localization Golgi Region 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Weizhen Bi
    • 1
  • Pedro J. Bonilla
    • 2
  • Kathryn V. Holmes
    • 3
  • Susan R. Weiss
    • 2
  • Julian L. Leibowitz
    • 1
  1. 1.Department of Pathology and Laboratory, Medicine and the Department of Microbiology and Molecular GeneticsUniversity of Texas Health Sciences CenterHoustonUSA
  2. 2.The Department of MicrobiologyUniversity of PennsylvaniaPhiladelphiaUSA
  3. 3.The Department of Pathology, Uniformed ServicesUniversity of the Health SciencesBethesdaUSA

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