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Comparative Study
. 2005 Dec 13;102(50):18153-8.
doi: 10.1073/pnas.0509201102. Epub 2005 Nov 30.

Human cytomegalovirus virion protein complex required for epithelial and endothelial cell tropism

Affiliations
Comparative Study

Human cytomegalovirus virion protein complex required for epithelial and endothelial cell tropism

Dai Wang et al. Proc Natl Acad Sci U S A. .

Abstract

Human cytomegalovirus replicates in many different cell types, including epithelial cells, endothelial cells, and fibroblasts. However, laboratory strains of the virus, many of which were developed as attenuated vaccine candidates by serial passage in fibroblasts, have lost the ability to infect epithelial and endothelial cells. Their growth is restricted primarily to fibroblasts, due to mutations in the UL131-UL128 locus. We now demonstrate that two products of this locus, pUL130 and pUL128, form a complex with gH and gL, but not gO. The AD169 laboratory strain, which lacks a functional UL131 protein, produces virions containing only the gH-gL-gO complex. An epithelial and endothelial cell tropic AD169 variant in which the UL131 ORF has been repaired, termed BADrUL131, produces virions that carry both gH-gL-gO and gH-gL-pUL128-pUL130 complexes. Antibodies against pUL130 and pUL128 block infection of epithelial and endothelial cells by BADrUL131 and the fusion-inducing factor X clinical human cytomegalovirus isolate but do not affect the efficiency with which fibroblasts are infected.

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Figures

Fig. 1.
Fig. 1.
pUL128 and pUL130 form a complex with gH. MRC-5 cells were infected with BADdlUL131-128, BADwt, or BADrUL131. Seventy-two hours later, cells were radioactively labeled for 1 h and chased for 20 or 120 min. Proteins were immunoprecipitated (IP) from cell lysates and analyzed by electrophoresis in an SDS-containing 12% polyacrylamide gel followed by autoradiography. Immunoprecipitations used anti-gB 7-17 (A), anti-gM IMP91-3/1 (B), anti-gH 14-4b (C), or anti-gH 14-4b, followed by anti-gO, anti-pUL130 3C5, or anti-pUL128 R551A (D). The positions at which marker proteins migrated are identified by their molecular weights (MW).
Fig. 2.
Fig. 2.
pUL128-pUL130 and gO form separate complexes with gH. MRC-5 cells were infected with BADdlUL131-128, BADwt, or BADrUL131. (A-C) Cells were radioactively labeled for 1 h and chased for 20 or 120 min beginning at 72 h postinfection. Proteins were immunoprecipitated (IP) from cell lysates and analyzed by electrophoresis in an SDS-containing 12% polyacrylamide gel followed by autoradiography. Immunoprecipitations used anti-gO (A), anti-pUL128 4B10 (B), or anti-pUL130 3E3 (C). (D) Displays combined immunoprecipitation and Western blot (WB) assays of pUL128-interacting proteins. Cells were lysed at 72 h postinfection, and extracts were subjected to immunoprecipitation with anti-pUL128 R551A antibody. The precipitated proteins were separated by electrophoresis in SDS-containing 12% polyacrylamide gels and analyzed by Western blot assay with anti-gH AP86, anti-pUL130 3C3, or anti-gO antibody. The positions at which marker proteins migrated are identified by their molecular weights (MW); antibody heavy (HC) and light chains (LC) are designated.
Fig. 3.
Fig. 3.
pUL128 and pUL130 are in virions. (A) BADwt and BADrUL131 virion proteins were analyzed by Western blot (WB) by using anti-gH AP86, anti-pUL130 3C5, or anti-pUL128 4B10 antibody. (B) Virion proteins were immunoprecipitated (IP) with anti-pUL128 R551A and analyzed by Western blot by using anti-gH AP86 or anti-pUL130 3C5 antibody. The positions at which marker proteins migrated are identified by their molecular weights (MW); antibody heavy chains (HC) are designated.
Fig. 4.
Fig. 4.
Characterization of gH-gL complexes. (A) Disulfide linkage of pUL128 with gH-gL. Purified BADrUL131 proteins in buffer with or without 2-mercaptoethanol (β-ME) were subjected to electrophoresis in an SDS-containing 12% (Left) or 4-20% (Right) polyacrylamide gel and analyzed by Western blot (WB) by using anti-pUL130 3C5 (Left) or anti-pUL128 4B10 (Right) antibody. (B) Comparison of complexes in BADwt and BADrUL131 virions. Virion proteins were separated by electrophoresis in SDS-containing, reducing or nonreducing, 8% polyacrylamide gels and analyzed by Western blot (WB) assay by using anti-gO (Left), anti-gH AP86 (Center), or anti-pUL128 4B10 (Right) antibody. *, monomeric gH; ♦ and • identify monomeric forms of gO. The positions at which marker proteins migrated are identified by their molecular weights (MW).
Fig. 5.
Fig. 5.
Neutralization of HCMV infectivity ARPE-19 epithelial cells, HUVEC endothelial cells, and MRC-5 fibroblasts. BADrUL131 (A)orBFXwt (B) were incubated with various concentrations of anti-pUL130 3C5 or 3E3 or anti-pUL128 R551A antibody, and residual infectivity was determined on different cell types.

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References

    1. Plachter, B., Sinzger, C. & Jahn, G. (1996) Adv. Virus Res. 46, 195-261. - PubMed
    1. Elek, S. D. & Stern, H. (1974) Lancet 1, 1-5. - PubMed
    1. Plotkin, S. A., Furukawa, T., Zygraich, N. & Huygelen, C. (1975) Infect. Immun. 12, 521-527. - PMC - PubMed
    1. Friedman, H. M., Macarak, E. J., MacGregor, R. R., Wolfe, J. & Kefalides, N. A. (1981) J. Infect. Dis. 143, 266-273. - PubMed
    1. Hahn, G., Revello, M. G., Patrone, M., Percivalle, E., Campanini, G., Sarasini, A., Wagner, M., Gallina, A., Milanesi, G., Koszinowski, U., et al. (2004) J. Virol. 78, 10023-10033. - PMC - PubMed

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