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(NAS Colloquium) Genetic Engineering of Viruses and Viral Vectors (1996) / Chapter Skim
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Human cytomegalovirus U53 impairs transport and maturation of major histocompatibility complex class I heavy chains
Pages 11327-11333

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From page 11327...
... In contrast to US2, expression of US3 does not destabilize heavy chains, but maturation and transport to the cell surface of MHC class I heavy chains are impaired. We show that a US3 gene product forms a complex with 52-microglobulin-associated class I heavy chains, which accumulate predominantly in the ER of US3+ cells.
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From page 11328...
... . Rabbit polyclonal antisera designated anti-HC, which reacts free MHC class I heavy chains, has been described (17~.
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From page 11329...
... U373 control cells or US3 + cells were metabolically radiolabeled for a 1-fur pulse and then chased for the indicated time (fur) in medium containing excess unlabeled amino acids.
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From page 11330...
... Thus, the predominant location of the impaired class I heavy chains is the ER in US3+ cells. Immunofluorescence microscopy of nonpermeabilized cells indicated that substantial amounts of class I heavy chains were on the surface of both US3 + cells and U373 control cells, while cell surface heavy chains were not detected in US2+ cells (Fig.
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From page 11331...
... digitonin lysis buffer or the standard lysis buffer containing sodium deoxycholate, sodium dodecyl sulfate, and Nonidet P-40. From the digitonin extracts, class I heavy chains could be recovered for each cell line using either anti-HC antibody or W6/32 antibody but not with the US3 antibody.
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From page 11332...
... DISCUSSION Herein we describe the effects of the product encoded by the HCMV US3 gene, a glycoprotein homologous to that encoded by US2, on expression of MHC class ~ products. By comparison of stably transfected cell lines that express either US2 or US3, we demonstrated that, in contrast to US2, expression of US3 does not cause rapid turnover of class I heavy chains.
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From page 11333...
... , and their attack on I3~-microglobulin-associated heavy chains is detected shortly after this association (21~. Furthermore, gpUS11 has been shown to cause destabilization of newly synthesized heavy chains upon completion of polypeptide synthesis by dislocating them from the ER to the cytosol, probably by reverse transport through the ER translocation complex (21~.
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Key Terms

  • mhc class
  • control cells
  • cell surface
  • lysis buffer
  • standard lysis

  • monoclonal antibody
  • heavy chain
  • immunofluorescence microscopy
  • gene product
  • amino acids

  • translocation complex
  • excess unlabeled
  • human cytomegalovirus
  • cell lines
  • unlabeled amino

  • containing excess
  • endoplasmic reticulum
  • unpublished results
  • digitonin lysis
  • medium containing

  • histocompatibility complex
  • major histocompatibility
  • metabolically radiolabeled
  • surface mhc
  • dodecyl sulfate

  • cell line
  • rapid turnover
  • surface proteins
  • indicated time
  • confocal immunofluorescence


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