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research article

The dynamics of yeast telomeres and silencing proteins through the cell cycle

Laroche, T  
•
Martin, S G
•
Tsai-Pflugfelder, M
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2000
Journal of structural biology

Genes integrated near the telomeres of budding yeast have a variegated pattern of gene repression that is mediated by the silent information regulatory proteins Sir2p, Sir3p, and Sir4p. Immunolocalization and fluorescence in situ hybridization (FISH) reveal 6-10 perinuclear foci in which silencing proteins and subtelomeric sequences colocalize, suggesting that these are sites of Sir-mediated repression. Telomeres lacking subtelomeric repeat elements and the silent mating locus, HML, also localize to the periphery of the nucleus. Conditions that disrupt telomere proximal repression disrupt the focal staining pattern of Sir proteins, but not necessarily the localization of telomeric DNA. To monitor the telomere-associated pools of heterochromatin-binding proteins (Sir and Rap1 proteins) during mitotic cell division, we have performed immunofluorescence and telomeric FISH on populations of yeast cells synchronously traversing the cell cycle. We observe a partial release of Rap1p from telomeres in late G2/M, although telomeres appear to stay clustered during G2-phase and throughout mitosis. A partial release of Sir3p and Sir4p during mitosis also occurs. This is not observed upon HU arrest, although other types of DNA damage cause a dramatic relocalization of Sir and Rap1 proteins. The observed cell cycle dynamics were confirmed by direct epifluorescence of a GFP-Rap1p fusion. Using live GFP fluorescence we show that the diffuse mitotic distribution of GFP-Rap1p is restored to the interphase pattern of foci in early G1-phase.

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Type
research article
DOI
10.1006/jsbi.2000.4240
Author(s)
Laroche, T  
Martin, S G
Tsai-Pflugfelder, M
Gasser, S M
Date Issued

2000

Published in
Journal of structural biology
Volume

129

Issue

2-3

Start page

159

End page

74

Subjects

Silent Information Regulator Proteins

•

Saccharomyces cerevisiae

Editorial or Peer reviewed

REVIEWED

Written at

OTHER

EPFL units
PTBIOP  
Available on Infoscience
February 19, 2009
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/35536
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