Dynamics of telomeric clusters relative to a single telomere. (A) 3D reconstruction of a yeast nucleus expressing Rap1-YFP, which labels all telomeres, and LacI-CFP, which recognizes a lacO array inserted on Tel14L (GA-2558). The larger view is rotated to allow visualization in the form of two other versions using Imaris software. The position of Tel14L (green) is scored relative to the Rap1 foci (red). (c) Tel14L colocalizing with a Rap1 focus, (ad) Tel14L adjacent to a Rap1 focus, (ap) Tel14L apart from Rap1 foci. Two-hundred-twenty nuclei reconstituted in 3D were analyzed, and in 70% of the nuclei Tel14L was either colocalizing or adjacent to the Rap1 focus. (B) Time-lapse imaging of the same yeast strain presented in A (GA-2558). 3D image stacks were taken at 30-sec intervals, deconvolved, and are shown in a 3D reconstitution. The top view is shown, and the juxtaposition of Tel14L to Rap1 was scored and indicated as in A. Continuous juxtaposition can be seen over 5 min.
