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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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Related in: MedlinePlus

UL135 Interacts with the WAVE2 Complex and Talin(A) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl, and HFFF were infected with HCMV strain Merlin or MerlinΔUL135. Samples were lysed 48 hr postinfection, and immunoprecipitation was performed for the V5 tag on UL135. Proteins were separated by SDS-PAGE, and western blot was performed for the indicated proteins.(B–I) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl (B, D, F, and H), and HFFF were infected with HCMV strain Merlin or MerlinΔUL135 or mock infected (C, E, G, and I). Samples were fixed and stained for UL135 (V5 antibody) and ABI1 (B and C), WAVE2 (D and E), or talin (H and I) 48 hr postinfection. Samples were lysed, and SDS-PAGE was performed 48 hr postinfection (F) or at the indicated time points (G) followed by western blot for the indicated proteins.
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fig4: UL135 Interacts with the WAVE2 Complex and Talin(A) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl, and HFFF were infected with HCMV strain Merlin or MerlinΔUL135. Samples were lysed 48 hr postinfection, and immunoprecipitation was performed for the V5 tag on UL135. Proteins were separated by SDS-PAGE, and western blot was performed for the indicated proteins.(B–I) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl (B, D, F, and H), and HFFF were infected with HCMV strain Merlin or MerlinΔUL135 or mock infected (C, E, G, and I). Samples were fixed and stained for UL135 (V5 antibody) and ABI1 (B and C), WAVE2 (D and E), or talin (H and I) 48 hr postinfection. Samples were lysed, and SDS-PAGE was performed 48 hr postinfection (F) or at the indicated time points (G) followed by western blot for the indicated proteins.

Mentions: Yeast two-hybrid assays and stable isotope labeling by amino acids in cell culture (SILAC) immunoprecipitation experiments were undertaken in order to identify pUL135-interacting partners. In the yeast two-hybrid screen, ABI1 and ABI2 were both identified in multiple clones. This finding was consolidated and expanded when SILAC immunoprecipitation experiments performed on pUL135-expressing fibroblasts identified WAVE2, ABI1, NAP1, CYFIP1, and talin-1 (Table S2). Significantly ABI1, ABI2, NAP1, CYFIP1, and WAVE2 itself are all components of the WAVE2 regulatory complex (WRC), which regulates the actin nucleator Arp2/3 (Takenawa and Suetsugu, 2007). The interaction with WAVE2, ABI1, ABI2, NAP1, CYFIP1, and talin-1 were validated in conventional immunoprecipitation experiments when pUL135 was expressed in both isolation and the context of HCMV infection (Figure 4A).


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 Interacts with the WAVE2 Complex and Talin(A) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl, and HFFF were infected with HCMV strain Merlin or MerlinΔUL135. Samples were lysed 48 hr postinfection, and immunoprecipitation was performed for the V5 tag on UL135. Proteins were separated by SDS-PAGE, and western blot was performed for the indicated proteins.(B–I) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl (B, D, F, and H), and HFFF were infected with HCMV strain Merlin or MerlinΔUL135 or mock infected (C, E, G, and I). Samples were fixed and stained for UL135 (V5 antibody) and ABI1 (B and C), WAVE2 (D and E), or talin (H and I) 48 hr postinfection. Samples were lysed, and SDS-PAGE was performed 48 hr postinfection (F) or at the indicated time points (G) followed by western blot for the indicated proteins.
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Related In: Results  -  Collection

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fig4: UL135 Interacts with the WAVE2 Complex and Talin(A) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl, and HFFF were infected with HCMV strain Merlin or MerlinΔUL135. Samples were lysed 48 hr postinfection, and immunoprecipitation was performed for the V5 tag on UL135. Proteins were separated by SDS-PAGE, and western blot was performed for the indicated proteins.(B–I) HFFF-hCAR were infected with RAd-UL135 or RAd-Ctrl (B, D, F, and H), and HFFF were infected with HCMV strain Merlin or MerlinΔUL135 or mock infected (C, E, G, and I). Samples were fixed and stained for UL135 (V5 antibody) and ABI1 (B and C), WAVE2 (D and E), or talin (H and I) 48 hr postinfection. Samples were lysed, and SDS-PAGE was performed 48 hr postinfection (F) or at the indicated time points (G) followed by western blot for the indicated proteins.
Mentions: Yeast two-hybrid assays and stable isotope labeling by amino acids in cell culture (SILAC) immunoprecipitation experiments were undertaken in order to identify pUL135-interacting partners. In the yeast two-hybrid screen, ABI1 and ABI2 were both identified in multiple clones. This finding was consolidated and expanded when SILAC immunoprecipitation experiments performed on pUL135-expressing fibroblasts identified WAVE2, ABI1, NAP1, CYFIP1, and talin-1 (Table S2). Significantly ABI1, ABI2, NAP1, CYFIP1, and WAVE2 itself are all components of the WAVE2 regulatory complex (WRC), which regulates the actin nucleator Arp2/3 (Takenawa and Suetsugu, 2007). The interaction with WAVE2, ABI1, ABI2, NAP1, CYFIP1, and talin-1 were validated in conventional immunoprecipitation experiments when pUL135 was expressed in both isolation and the context of HCMV infection (Figure 4A).

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