spacer gif spacer gif spacer gif spacer gif spacer gif
 QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

First published online October 27, 2005
doi: 10.1242/10.1242/jcs.02612


Journal of Cell Science 118, 4985-4994 (2005)
Published by The Company of Biologists 2005
This Article
Right arrow Summary Freely available
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Trelles-Sticken, E.
Right arrow Articles by de La Roche Saint-André, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Trelles-Sticken, E.
Right arrow Articles by de La Roche Saint-André, C.
Social Bookmarking
 Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

Set1- and Clb5-deficiencies disclose the differential regulation of centromere and telomere dynamics in Saccharomyces cerevisiae meiosis

Edgar Trelles-Sticken1,*, Sandrine Bonfils2, Julie Sollier2, Vincent Géli2, Harry Scherthan1,{ddagger},§ and Christophe de La Roche Saint-André2,§

1 MPI for Molecular Genetics, Ihnestr. 73, 14195 Berlin, Germany
2 LISM, CNRS, 31 chemin Joseph Aiguier, 13402 Marseille, Cedex 20, France



View larger version (15K):

[in a new window]
 
Fig. 1. Analysis of centromere and telomere dynamics during meiosis in wild-type diploid SK1 cells. Frequencies of nuclei displaying a single centromere FISH signal (% cen cluster), of nuclei with two to eight perinuclear vegetative telomere FISH signals (% veg tel cluster), of nuclei containing a single telomere FISH signal (% bouquet) and of nuclei with paired signals with a cosmid probe hybridizing to an internal region of the left arm of chromosome XI (% pairing), at different time points (minutes) after transfer into sporulation medium (SPM). Top: FACS profiles corresponding to the same time points.

 


View larger version (13K):

[in a new window]
 
Fig. 2. Centromere and telomere dynamics during meiosis in clb5{Delta}. Frequencies of nuclei displaying a single centromere FISH signal (% cen cluster), of nuclei with two to eight perinuclear vegetative telomere FISH signals (% veg tel cluster) or with a single telomere signal (% bouquet) in wild-type and clb5{Delta} strains, at the indicated time points after transfer into sporulation medium. Top: wild-type and clb5{Delta} FACS profiles aligned to the corresponding time points.

 


View larger version (19K):

[in a new window]
 
Fig. 3. Centromere and telomere dynamics during meiosis in ime2{Delta}. Frequencies of ime2{Delta} nuclei displaying a single centromere FISH signal (% cen cluster), with two to eight perinuclear vegetative telomere FISH signals (% veg tel cluster) or with a single telomere FISH signal (% bouquet), at the indicated time points after transfer into sporulation medium. Top: wild-type and ime2{Delta} FACS profiles aligned to the corresponding time points.

 


View larger version (13K):

[in a new window]
 
Fig. 4. Loss of Set1 limits chromosome dynamics during the meiotic prophase. Frequencies of nuclei displaying a single centromere FISH signal (% cen cluster), with two to eight perinuclear vegetative telomere FISH signals (% veg tel cluster) or with a single telomere FISH signal (% bouquet) in wild-type and set1{Delta} strains, at the indicated time points after transfer into sporulation medium. Solid and dotted lines: data from two independent time course experiments. Top: wild-type and set1{Delta} FACS profiles from the experiment shown by the solid line, aligned to the corresponding time points.

 


View larger version (13K):

[in a new window]
 
Fig. 5. Epistasis analysis of the set1{Delta} and clb5{Delta} mutations. Frequencies of nuclei displaying a single centromere FISH signal (% cen cluster), with two to eight perinuclear telomere FISH signals (% veg tel cluster) or with a single telomere FISH signal cluster (% bouquet) in clb5{Delta} and set1{Delta} clb5{Delta} strains, at the indicated time points after transfer into sporulation medium. Top: clb5{Delta} and set1{Delta} clb5{Delta} FACS profiles aligned to the corresponding time points.

 


View larger version (22K):

[in a new window]
 
Fig. 6. Analysis of synapsis and pairing in set1{Delta}, clb5{Delta} and set1{Delta} clb5{Delta} cells. (A) Frequencies of nuclei with paired cosmid signals during meiotic time courses; wild-type and set1{Delta} strains (left graphs), or the wild-type, clb5{Delta} and set1{Delta} clb5{Delta} strains (right graphs). Top graphs: frequencies of nuclei displaying only a single signal due to pairing of a cosmid probe corresponding to a telomeric region of the chromosome III left arm (cos m; 37). Bottom graphs: frequencies of nuclei displaying paired signals with a cosmid probe hybridizing an internal region of the chromosome XI left arm (cos f; 37). (B) Representative images of Zip1-positive nuclei at 330 minutes (top) or 270 minutes (bottom) after induction of meiosis. WT, wild-type nuclei displaying thread-like Zip1 signals (green); set1{Delta} nuclei displaying Zip1 speckles only. In set1{Delta} clb5{Delta} this is only the case of a few nuclei in the culture, whereas in clb5{Delta} a few nuclei display a Zip1 polycomplex (bright green oblong). Bar, 5 µm.

 


View larger version (13K):

[in a new window]
 
Fig. 7. Role of Mec1 in restricting chromosome reorganization in set1{Delta} cells. Frequencies of nuclei displaying a single centromere FISH signal (% cen cluster), two to eight perinuclear telomere FISH signals (% tel cluster) or a single telomere FISH signal cluster (% bouquet) in wild-type and set1{Delta} mec1-1 strains, at the indicated time points after transfer into sporulation medium. Top: wild-type and set1{Delta} mec1-1 FACS profiles aligned to the corresponding time points.

 


View larger version (32K):

[in a new window]
 
Fig. 8. Localization of Ndj1 and Rap1 is normal in set1{Delta} meiotic nuclei. (A) Frequency of nuclei from wild-type and set1{Delta} diploid (HA)-tagged Ndj1 strains displaying Ndj1-HA signal at the indicated time point in sporulation medium. (B) Colocalization of Ndj1-HA immunofluorescence signals (red) and XY' telo-FISH signals (green) in representative wild-type and set1{Delta} nuclei. Bar, 5 µm. (C) Immunolocalization of Rap1 (middle panels) and Ndj1-HA (right panels) in mildly spread wild-type (top panels) and set1{Delta} (bottom panels) nuclei. Single immunofluorescence channels are shown in gray scale for better sensitivity. Left panels: merged color images of immunofluorescence (green: Rap1; red: Ndj1-HA). A Ndj1-HA-positive spot with no detectable Rap1 signal is indicated (white arrowhead). Bar, 5 µm.

 


View larger version (11K):

[in a new window]
 
Fig. 9. Scheme showing the sequential steps of centromere and telomere redistribution that occur during transit from vegetative nuclear architecture into meiotic prophase. It is based on observations made in this and earlier work (see text). In the premeiotic nucleus, telomeres (red) form several perinuclear clusters and all centromeres (blue) locate tightly near the spindle pole body (SPB). Induction of meiosis leads to the dissolution of the centromere cluster and perinuclear telomere clusters. The dispersion of centromeres and telomeres differ relative to their dependence on premeiotic S phase. Telomeres cluster transiently near the SPB (bouquet stage) before scattering again over the periphery of the nucleus. The steps where centromere and telomere behavior are affected by the loss of Clb5 and Set1 are indicated by blue and red T-bars, respectively. The absence of Clb5 affects only the movements of telomeres with a severe impairment of vegetative telomere cluster dissolution, a delayed occurrence of meiotic telomere clustering and a persistence of meiotic telomere clustering. The lack of Set1, on the other hand, induces defects in centromere and telomere dispersion from their respective vegetative clusters and blocks meiotic telomere cluster formation. Accordingly, the thin T-bars represent a leaky effect which can be bypassed by some cells to reach the next stage of nuclear topology, whereas the bold T-bars represent a block that induces a cell to remain with a topological pattern of the previous stage, while prophase I may still be progressing. Note that the relative duration of the transitions is not drawn to scale.

 

Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?




© The Company of Biologists Ltd 2005