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


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

doi: 10.1242/10.1242/jcs.00299

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 Related articles in JCS
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 Wachtler, V.
Right arrow Articles by Balasubramanian, M. K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wachtler, V.
Right arrow Articles by Balasubramanian, M. K.
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?

Sterol-rich plasma membrane domains in the fission yeast Schizosaccharomyces pombe

Volker Wachtler, Srividya Rajagopalan and Mohan K. Balasubramanian*

Cell Division Laboratory, Temasek Life Sciences Laboratory, 1 Research Link, The National University of Singapore, Singapore 117604



View larger version (70K):

[in a new window]
  		Fig. 1. Sterols localise to distinct sites of the plasma membrane. (A) Sterol localisation detected by filipin staining in wild-type cells. (B) Frequency of filipin staining patterns denoted in A in an exponentially growing wild-type culture.

 


View larger version (66K):

[in a new window]
  		Fig. 2. Sterol localisation correlates with sites of active cell growth and cytokinesis. Sterol localisation detected by filipin staining in (A) nitrogen-starved cells, (B) cells released from nitrogen starvation for 80 minutes, (C) cdc10-V50 cells at 36°C, (E) glucose-starved cells, (F) cells grown in medium with glucose, (G) cdc25-22 cells at 36°C, (H) orb1-13 cells at 36°C, (I) a branched TBZ-treated cdc10-V50 cell, (J) mating cells, (K) ascospores and (L) mid1-18 cells at 36°C. (D) Localisation of CHD-GFP in cdc10-V50 cells.

 


View larger version (73K):

[in a new window]
  		Fig. 3. Sterol localisation to the middle of the cell during mitosis and cytokinesis. cdc25-22 hht2GFP rlc1GFP cells were synchronised and released to the permissive temperature (18°C). Samples were taken at the indicated time points after release, stained with filipin and observed for GFP and filipin fluorescence.

 


View larger version (58K):

[in a new window]
  		Fig. 4. Sterol localisation to the middle of the cell requires a functional secretory pathway but not an intact F-actin or microtubule cytoskeleton. (A,C) cdc25-22 cells were synchronised and released to the permissive temperature (24°C). Cells were released into medium containing (A) 100 µM LatA or 1% (v/v) DMSO as solvent control, or (C) 100 µM BFA or 1% (v/v) EtOH as solvent control. Samples were taken 60 minutes after release and stained with filipin. (B,D) Sterol localisation detected by filipin staining in (B) nda3-KM311 cells at 18°C and cells overexpressing mad2+ under the nmt1-promotor, and (D) cdc7-24 cells at 36°C. Arrows indicate strong medial staining in A and faint medial staining in C and D. Arowheads indicate additional asymmetric patches of sterol-rich membrane.

 


View larger version (56K):

[in a new window]
  		Fig. 5. Structural alterations of sterol-rich membrane domains affect cytokinesis in S. pombe. (A,B) cdc4GFP cells were grown for 1 hour in medium containing 5 µg/ml filipin or 0.1% (v/v) DMSO as solvent control. Arrow indicates mis-shapen Cdc4-GFP ring; arrowhead indicates Cdc4-GFP spots. (C) cdc4GFP cells overexpressing C-4 sterol methyl oxidase under the nmt1-promotor. (D,E) Quantitation of localisation patterns. Bars in D represent filipin-treated cells (grey) and DMSO-treated cells (white); bars in E represent cells overexpressing C-4 sterol methyl oxidase (dark grey), control cells with empty vector (light grey) and control cells in 15 µM thiamine (white).

 


View larger version (57K):

[in a new window]
  		Fig. 6. Structural alterations of sterol-rich membrane domains compromise the stability of a colocalising plasma membrane protein. (A,B) bgs4GFP cells were grown for 1 hour in medium containing 5 µg/ml filipin or 0.1% (v/v) DMSO as solvent control. (C) Equal amounts (11.5 µg each) of total protein from filipin- or DMSO-treated cells were analysed by immunoblotting and probed with anti-GFP antibodies. The signal strength was quantified and plotted as relative units.

 

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 2003