Identfication of protein domains involved in nuclear localization and dimerization of TIM.

<p><b>A</b>) Schematic diagram of the V5-tagged long (l-full length) and short TIM proteins (s), as well as GFP-tagged truncated versions, including the position of the putative nuclear localization signals (red) and coiled-coil domains (yellow). Subcellular localization, as observed during the experiments is represented by N (nuclear) or C (cytoplasmic). <b>B</b>) Representative examples of the subcellular distribution patterns of the various tagged TIM proteins in COS7 cells, as detected by GFP fluorescence or staining with anti-V5 antibodies. Note that TIM(1–309)-GFP appears to be subject to accelerated proteolytic degradation, as it is expressed at lower level and shows signs of accumulation at the aggresome (arrow), a cytoplasmic structure involved in 26S proteasome degradation. <b>C</b>) Representative examples of COS7 cells transiently expressing truncated TIM(1–1079)-GFP or TIM(1–309)-GFP proteins (green) together with l-TIM-V5 (blue). Arrows show truncated TIM-GFP proteins that are translocated to the nucleus in presence of l-TIM-V5.