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PMID: 15539461 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Finding new components of the target of rapamycin (TOR) signaling network through chemical genetics and proteome chips.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 101 ·No. 47 ·2004-11-23 ·Pages 16594-9

Huang J, Zhu H, Haggarty SJ, Spring DR, Hwang H, Jin F, Snyder M, Schreiber SL

Abstract

The TOR (target of rapamycin) proteins play important roles in nutrient signaling in eukaryotic cells. Rapamycin treatment induces a state reminiscent of the nutrient starvation response, often resulting in growth inhibition. Using a chemical genetic modifier screen, we identified two classes of small molecules, small-molecule inhibitors of rapamycin (SMIRs) and small-molecule enhancers of rapamycin (SMERs), that suppress and augment, respectively, rapamycin's effect in the yeast Saccharomyces cerevisiae. Probing proteome chips with biotinylated SMIRs revealed putative intracellular target proteins, including Tep1p, a homolog of the mammalian PTEN (phosphatase and tensin homologue deleted on chromosome 10) tumor suppressor, and Ybr077cp (Nir1p), a protein of previously unknown function that we show to be a component of the TOR signaling network. Both SMIR target proteins are associated with PI(3,4)P2, suggesting a mechanism of regulation of the TOR pathway involving phosphatidylinositides. Our results illustrate the combined use of chemical genetics and proteomics in biological discovery and map a path for creating useful therapeutics for treating human diseases involving the TOR pathway, such as diabetes and cancer.

MeSH Terms
Humans Jurkat Cells Models, Biological Protein Array Analysis Protein Kinases/genetics,metabolism Proteomics Saccharomyces cerevisiae/drug effects,genetics,metabolism Saccharomyces cerevisiae Proteins/genetics,metabolism Signal Transduction Sirolimus/pharmacology TOR Serine-Threonine Kinases
Chemicals
Saccharomyces cerevisiae Proteins Protein Kinases MTOR protein, human TOR Serine-Threonine Kinases Sirolimus
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Huang Jing
Howard Hughes Medical Institute, Harvard Institute of Chemistry and Cell Biology, and Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA. [email protected]
Zhu Heng
Haggarty Stephen J
Spring David R
Hwang Heejun
Jin Fulai
Snyder Michael
Schreiber Stuart L
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0407117101
Email
[email protected]
Published
2004-11-23
Epub
2004-00-11
Pages
16594-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC527135
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.0407117101 PMC Full Text View journal details
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