BIOMEDICAL IMAGING GROUP (BIG)
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Controlled Exchange of Chromosomal Arms Reveals Principles Driving Telomere Interactions in Yeast

H. Schober, V. Kalck, M.A. Vega-Palas, G. Van Houwe, D. Sage, M. Unser, M.R. Gartenberg, S.M. Gasser

Genome Research, vol. 18, no. 2, pp. 261–271, February 2008.

The 32 telomeres in the budding yeast genome cluster in three to seven perinuclear foci. Although individual telomeres and telomeric foci are in constant motion, preferential juxtaposition of some telomeres has been scored. To examine the principles that guide such long-range interactions, we differentially tagged pairs of chromosome ends and developed an automated three-dimensional measuring tool that determines distances between two telomeres. In yeast, all chromosomal ends terminate in TG1-3 and middle repetitive elements, yet subgroups of telomeres also share extensive homology in subtelomeric coding domains. We find that up to 21 kb of >90% sequence identity does not promote telomere pairing in interphase cells. To test whether unique sequence elements, arm length, or chromosome territories influence juxtaposition, we reciprocally swapped terminal domains or entire chromosomal arms from one chromosome to another. We find that the distal 10 kb of Tel6R promotes interaction with Tel6L, yet only when the two telomeres are present on the same chromosome. By manipulating the length and sequence composition of the right arm of chr 5, we confirm that contact between telomeres on opposite chromatid arms of equal length is favored. These results can be explained by the polarized Rabl arrangement of yeast centromeres and telomeres, which promote to telomere pairing by allowing contact between chromosome arms of equal length in anaphase.

Supplementary data

  • Supplementary Figure Legends (DOC file) (60 kb). Legends, captions, and additional references.
  • Supplementary Materials and Methods (DOC file) (38 kb). Targeted recombination and additional reference.
  • Supplementary Table (DOC file) (121 kb). Strains used in this study.
  • Supplementary Figure 1 (PDF file) (1126 kb). Demonstration of the method for 3D distance determination.
  • Supplementary Figure 2 (PDF file) (778 kb). Data examples from the SpotDistance plug-in.
  • Supplementary Figure 3 (PDF file) (976 kb). Telomeric organization in S. cerevisiæ.
  • Supplementary Figure 4 (PDF file) (90 kb). The Tel6R pairing element works only in cis.
  • Supplementary Figure 5 (PDF file) (517 kb). Detailed description of the chromosome swap technique.
  • Supplementary Figure 6 (PDF file) (863 kb). Perinuclear anchoring does not correlate with clustering.
  • Supplementary Figure 7 (PDF file) (1524 kb). Annotations sites for transcriptions factors in the subtelomeric regions of telomeres 6R, 5R, 6L and 6RΔ10kb.
  • Supplementary Movie 1 (MOV file) (686 kb). A full chip Rap1-GFP 4D acquisition performed as described in Materials and Methods is displayed as a Z-projection. Total number of frames: 271 or 90 min of life imaging with a time frame of 20 s. The image stacks were deconvolved using the Huygens© software.
  • Supplementary Movie 2 (MOV file) (215 kb). SIR3-GFP time series displayed as a full rotation of a kymograph. Total time is 60 min.
  • Supplementary Movie 3 (MOV file) (249 Mb). Rap1-YFP (in red) and telomere 14L (in green) time-lapse; 30 sec intervals. The image stacks were deconvolved using the Huygens© software. Changing gray levels due to bleaching were adjusted using Imaris (Bitplane).
  • Supplementary Movie 4 (MOV file) (66 kb). Rap1-YFP (in red) and telomere 14L (in green) time-lapse; 30 sec intervals. The image stacks were deconvolved using the Huygens© software. Changing gray levels due to bleaching were adjusted using Imaris (Bitplane). 

@ARTICLE(http://bigwww.epfl.ch/publications/schober0801.html,
AUTHOR="Schober, H. and Kalck, V. and Vega-Palas, M.A. and Van Houwe, G.
	and Sage, D. and Unser, M. and Gartenberg, M.R. and Gasser, S.M.",
TITLE="Controlled Exchange of Chromosomal Arms Reveals Principles
	Driving Telomere Interactions in Yeast",
JOURNAL="Genome Research",
YEAR="2008",
volume="18",
number="2",
pages="261--271",
month="February",
note="")

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