This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
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 arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Sturgill, T. W.
Right arrow Articles by Hall, M. N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Sturgill, T. W.
Right arrow Articles by Hall, M. N.

 Previous Article  |  Next Article 

Eukaryotic Cell, October 2008, p. 1819-1830, Vol. 7, No. 10
1535-9778/08/$08.00+0     doi:10.1128/EC.00088-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

TOR1 and TOR2 Have Distinct Locations in Live Cells{triangledown} ,{dagger}

Thomas W. Sturgill,1* Adiel Cohen,2 Melanie Diefenbacher,2 Mark Trautwein,2 Dietmar E. Martin,2 and Michael N. Hall2

Department of Pharmacology, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908,1 Biozentrum, University of Basel, CH-4056 Basel, Switzerland2

Received 11 March 2008/ Accepted 9 July 2008

TOR is a structurally and functionally conserved Ser/Thr kinase found in two multiprotein complexes that regulate many cellular processes to control cell growth. Although extensively studied, the localization of TOR is still ambiguous, possibly because endogenous TOR in live cells has not been examined. Here, we examined the localization of green fluorescent protein (GFP) tagged, endogenous TOR1 and TOR2 in live S. cerevisiae cells. A DNA cassette encoding three copies of green fluorescent protein (3XGFP) was inserted in the TOR1 gene (at codon D330) or the TOR2 gene (at codon N321). The TORs were tagged internally because TOR1 or TOR2 tagged at the N or C terminus was not functional. The TOR1D330-3XGFP strain was not hypersensitive to rapamycin, was not cold sensitive, and was not resistant to manganese toxicity caused by the loss of Pmr1, all indications that TOR1-3XGFP was expressed and functional. TOR2-3XGFP was functional, as TOR2 is an essential gene and TOR2N321-3XGFP haploid cells were viable. Thus, TOR1 and TOR2 retain function after the insertion of 748 amino acids in a variable region of their noncatalytic domain. The localization patterns of TOR1-3XGFP and TOR2-3XGFP were documented by imaging of live cells. TOR1-3XGFP was diffusely cytoplasmic and concentrated near the vacuolar membrane. The TOR2-3XGFP signal was cytoplasmic but predominately in dots at the plasma membrane. Thus, TOR1 and TOR2 have distinct localization patterns, consistent with the regulation of cellular processes as part of two different complexes.

* Corresponding author. Mailing address: Department of Pharmacology, Box 800735, Room 5045, Jordan Hall, University of Virginia Health Sciences Center, Charlottesville, VA 22908. Phone: (434) 924-1919. Fax: (434) 924-5207. E-mail: Thomas_Sturgill{at}virginia.edu

{triangledown} Published ahead of print on 22 August 2008.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

Eukaryotic Cell, October 2008, p. 1819-1830, Vol. 7, No. 10
1535-9778/08/$08.00+0     doi:10.1128/EC.00088-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»