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HCMV pUL135 remodels the actin cytoskeleton to impair immune recognition of infected cells.

Stanton RJ, Prod'homme V, Purbhoo MA, Moore M, Aicheler RJ, Heinzmann M, Bailer SM, Haas J, Antrobus R, Weekes MP, Lehner PJ, Vojtesek B, Miners KL, Man S, Wilkie GS, Davison AJ, Wang EC, Tomasec P, Wilkinson GW - Cell Host Microbe (2014)

Bottom Line: Without immune pressure, laboratory-adapted HCMV strains have undergone genetic alterations.Among these, the deletion of the UL/b' domain is associated with loss of virulence.An independent interaction between pUL135 and talin disrupted cell contacts with the extracellular matrix.

View Article: PubMed Central - PubMed

Affiliation: Institute of Infection & Immunity, Cardiff University School of Medicine, Heath Park, Cardiff CF14 4XN, UK. Electronic address: stantonrj@cf.ac.uk.

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UL135 Modifies Actin Structure(A–D) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl (A and C), or HFFF were infected with HCMV strain Merlin or MerlinΔUL135 or mock infected (B and D). Samples were stained 48 hr postinfection for UL135 (V5 antibody) and actin (phalloidin; A and B). Following infection, samples were incubated with the indicated doses of LatA (LatA; C and D).NK cell degranulation assays were performed 48 hr postinfection with IFN-α-activated PBMC. Results are mean ± SD and are representative of three independent experiments. Two-way ANOVA test with post tests were performed between RAd-Ctrl and RAd-UL135 (C) or Merlin and MerlinΔUL135 (D). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.(E) Samples treated as in (C and D) were stained for actin (phalloidin).
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fig3: UL135 Modifies Actin Structure(A–D) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl (A and C), or HFFF were infected with HCMV strain Merlin or MerlinΔUL135 or mock infected (B and D). Samples were stained 48 hr postinfection for UL135 (V5 antibody) and actin (phalloidin; A and B). Following infection, samples were incubated with the indicated doses of LatA (LatA; C and D).NK cell degranulation assays were performed 48 hr postinfection with IFN-α-activated PBMC. Results are mean ± SD and are representative of three independent experiments. Two-way ANOVA test with post tests were performed between RAd-Ctrl and RAd-UL135 (C) or Merlin and MerlinΔUL135 (D). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.(E) Samples treated as in (C and D) were stained for actin (phalloidin).

Mentions: Expression of pUL135 in fibroblasts consistently induced dramatic changes in cell morphology, and focal adhesions and cell projections are lost as cells are rounded up (Figures 2D and S2C). Even in the context of productive infection, pUL135 clearly influenced the HCMV-induced cytopathic effect. In contrast to parental virus, cells infected with MerlinΔUL135 were clearly “flatter” and more spread out, which is indicative of enhanced adherence (Figures 2E and S2D). The actin cytoskeleton was stained with phalloidin; pUL135 expression resulted in elimination of F-actin stress fibers from the cell center, whereas the cortical actin matrix underlying the plasma membrane appeared reinforced and colocalized with pUL135 (Figures 3A and S3A). During productive HCMV infection, F-actin was lost from the center of infected cells, and UL135 colocalized with cortical actin. However, in the absence of pUL135, a substantial proportion of F-actin remained in the center of infected cells (Figures 3B and S3B). In contrast, microtubules and intermediate filaments were not overtly displaced by pUL135 (Figures S3C–S3F).


HCMV pUL135 remodels the actin cytoskeleton to impair immune recognition of infected cells.

Stanton RJ, Prod'homme V, Purbhoo MA, Moore M, Aicheler RJ, Heinzmann M, Bailer SM, Haas J, Antrobus R, Weekes MP, Lehner PJ, Vojtesek B, Miners KL, Man S, Wilkie GS, Davison AJ, Wang EC, Tomasec P, Wilkinson GW - Cell Host Microbe (2014)

UL135 Modifies Actin Structure(A–D) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl (A and C), or HFFF were infected with HCMV strain Merlin or MerlinΔUL135 or mock infected (B and D). Samples were stained 48 hr postinfection for UL135 (V5 antibody) and actin (phalloidin; A and B). Following infection, samples were incubated with the indicated doses of LatA (LatA; C and D).NK cell degranulation assays were performed 48 hr postinfection with IFN-α-activated PBMC. Results are mean ± SD and are representative of three independent experiments. Two-way ANOVA test with post tests were performed between RAd-Ctrl and RAd-UL135 (C) or Merlin and MerlinΔUL135 (D). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.(E) Samples treated as in (C and D) were stained for actin (phalloidin).
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fig3: UL135 Modifies Actin Structure(A–D) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl (A and C), or HFFF were infected with HCMV strain Merlin or MerlinΔUL135 or mock infected (B and D). Samples were stained 48 hr postinfection for UL135 (V5 antibody) and actin (phalloidin; A and B). Following infection, samples were incubated with the indicated doses of LatA (LatA; C and D).NK cell degranulation assays were performed 48 hr postinfection with IFN-α-activated PBMC. Results are mean ± SD and are representative of three independent experiments. Two-way ANOVA test with post tests were performed between RAd-Ctrl and RAd-UL135 (C) or Merlin and MerlinΔUL135 (D). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.(E) Samples treated as in (C and D) were stained for actin (phalloidin).
Mentions: Expression of pUL135 in fibroblasts consistently induced dramatic changes in cell morphology, and focal adhesions and cell projections are lost as cells are rounded up (Figures 2D and S2C). Even in the context of productive infection, pUL135 clearly influenced the HCMV-induced cytopathic effect. In contrast to parental virus, cells infected with MerlinΔUL135 were clearly “flatter” and more spread out, which is indicative of enhanced adherence (Figures 2E and S2D). The actin cytoskeleton was stained with phalloidin; pUL135 expression resulted in elimination of F-actin stress fibers from the cell center, whereas the cortical actin matrix underlying the plasma membrane appeared reinforced and colocalized with pUL135 (Figures 3A and S3A). During productive HCMV infection, F-actin was lost from the center of infected cells, and UL135 colocalized with cortical actin. However, in the absence of pUL135, a substantial proportion of F-actin remained in the center of infected cells (Figures 3B and S3B). In contrast, microtubules and intermediate filaments were not overtly displaced by pUL135 (Figures S3C–S3F).

Bottom Line: Without immune pressure, laboratory-adapted HCMV strains have undergone genetic alterations.Among these, the deletion of the UL/b' domain is associated with loss of virulence.An independent interaction between pUL135 and talin disrupted cell contacts with the extracellular matrix.

View Article: PubMed Central - PubMed

Affiliation: Institute of Infection & Immunity, Cardiff University School of Medicine, Heath Park, Cardiff CF14 4XN, UK. Electronic address: stantonrj@cf.ac.uk.

Show MeSH
Related in: MedlinePlus