FIGURE 4:

VPS4 residues required for ESCRT-III interaction are highly conserved in MITD1 and important for MITD1 midbody localization and ESCRT-III interaction. (A) Amino acid sequences of MIT domains of human MITD1 and VPS4A/B were aligned using ClustalW2. An asterisk indicates fully conserved residues. Colon indicates conservation between groups of strongly similar properties. Period indicates conservation between groups of weakly similar properties. (B–D) HEK293 cells were transfected with the indicated constructs and immunoprecipitated (IP) with anti-Myc antibodies. Precipitates were then immunoblotted (IB) with anti–HA- and anti–Myc-epitope antibodies. (E) HeLa cells were transfected with the indicated Myc-MITD1 constructs and immunostained for Myc epitope (red) and CHMP2A (green). Insets are enlarged images of midbody regions. HC, IgG heavy chain; LC, IgG light chain; WT, wild type. Bars, 2 μm.