Human Cytomegalovirus UL29/28 Protein Interacts with Components of the NuRD Complex Which Promote Accumulation of Immediate-Early RNA
Figure 2
HCMV pUL29/28 and pUL38 specifically bind to constituents of the NuRD complex.
Fibroblasts were infected at a multiplicity of 3 pfu/cell using BADinUL29F (inUL29F) virus, and whole cell lysates were prepared 24 h later. (A) HDAC1 and MTA2 coprecipitate with pUL29/28 and pUL38. Immunoprecipitations (IP) were performed using antibodies to MTA2 or endogenous HDAC1. Western blot (WB) analysis employed an anti-FLAG (pUL29/28) or pUL38 antibodies, and included analysis of lysates as a control. An uninfected control sample was also included. (B) pUL29/28 coprecipitates HDAC1, MTA2 and pUL38. Left panel: The immunoprecipitation was carried out using an anti-FLAG antibody (pUL29/28) or an isotype-specific anti-myc control antibody followed by Western blot analysis using antibodies to pUL38 and MTA2. Right panel: The experiment was repeated using antibody to HDAC1. (C) pUL29/28 interacts with the NuRD and Sin3 complexes. Left panel: Immunoprecipitation from whole cell lysates from uninfected or BADinUL29F (inUL29F) infected cells using antibodies to mSin3A and MTA2. Western blot analysis with antibodies against FLAG (pUL29/28), pUL38, MTA2 or mSin3A. Right panel: No evidence for interaction of pUL29/28 with tuberous sclerosis protein 2 (TSC2). Immunoprecipitations used antibodies against TSC2 and MTA2 and Western blots were performed with antibodies to FLAG (pUL29/28).