Probing the membrane environment of the TOR kinases reveals functional interactions between TORC1, actin, and membrane trafficking in Saccharomyces cerevisiae

Sofia Aronova, Karen Wedaman, Scott Anderson, John Yates, Ted Powers

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

The TOR kinases are regulators of growth in eukaryotic cells that assemble into two distinct protein complexes, TORC1 and TORC2, where TORC1 is inhibited by the antibiotic rapamycin. Present models favor a view wherein TORC1 regulates cell mass accumulation, and TORC2 regulates spatial aspects of growth, including organization of the actin cytoskeleton. Here, we demonstrate that in yeast both TORC1 and TORC2 fractionate with a novel form of detergent-resistant membranes that are distinct from detergent-resistant plasma membrane "rafts." Proteomic analysis of these TOR-associated membranes revealed the presence of regulators of endocytosis and the actin cytoskeleton. Genetic analyses revealed a significant number of interactions between these components and TORC1, demonstrating a functional link between TORC1 and actin/endocytosis-related genes. Moreover, we found that inhibition of TORC1 by rapamycin 1) disrupted actin polarization, 2) delayed actin repolarization after glucose starvation, and 3) delayed accumulation of lucifer yellow within the vacuole. By combining our genetic results with database mining, we constructed a map of interactions that led to the identification of additional genetic interactions between TORC1 and components involved in membrane trafficking. Together, these results reveal the broad scope of cellular processes influenced by TORC1, and they underscore the functional overlap between TORC1 and TORC2.

Original languageEnglish (US)
Pages (from-to)2779-2794
Number of pages16
JournalMolecular Biology of the Cell
Volume18
Issue number8
DOIs
StatePublished - Aug 2007

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TOR Serine-Threonine Kinases
Saccharomyces cerevisiae
Actins
Membranes
Sirolimus
Endocytosis
Actin Cytoskeleton
Detergents
mechanistic target of rapamycin complex 1
Eukaryotic Cells
Growth
Starvation
Vacuoles
Proteomics
Yeasts

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Probing the membrane environment of the TOR kinases reveals functional interactions between TORC1, actin, and membrane trafficking in Saccharomyces cerevisiae. / Aronova, Sofia; Wedaman, Karen; Anderson, Scott; Yates, John; Powers, Ted.

In: Molecular Biology of the Cell, Vol. 18, No. 8, 08.2007, p. 2779-2794.

Research output: Contribution to journalArticle

Aronova, Sofia ; Wedaman, Karen ; Anderson, Scott ; Yates, John ; Powers, Ted. / Probing the membrane environment of the TOR kinases reveals functional interactions between TORC1, actin, and membrane trafficking in Saccharomyces cerevisiae. In: Molecular Biology of the Cell. 2007 ; Vol. 18, No. 8. pp. 2779-2794.
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