FIGURE 6.

GCP4 and GCP5 interact directly within γTuRCs. A, localization of GCP4-GFP (green) and GCP5-mCherry (red) in cells overexpressing Nlp. B, GFP lifetime distribution of GCP4-GFP with or without co-expression of GCP5-mCherry. C, GFP lifetime distribution of GCP4-GFP with or without co-expression of GCP4-mCherry. D, Western blot analysis of GCP4 and GCP5 after cross-linking of purified γTuRCs in vitro. At low concentration of cross-linker, a specific band containing GCP4 (76 kDa) and GCP5 (118 kDa) appears at the size of GCP6 (used here as a marker of 200 kDa), corresponding to the size of the heterodimer. An additional band containing GCP4 was also observed around 130 kDa (*).