Abstract
Genome-wide copy number analyses of human cancers identified a frequent 5p13 amplification in several solid tumour types, including lung (56%), ovarian (38%), breast (32%), prostate (37%) and melanoma (32%). Here, using integrative analysis of a genomic profile of the region, we identify a Golgi protein, GOLPH3, as a candidate targeted for amplification. Gain- and loss-of-function studies in vitro and in vivo validated GOLPH3 as a potent oncogene. Physically, GOLPH3 localizes to the trans-Golgi network and interacts with components of the retromer complex, which in yeast has been linked to target of rapamycin (TOR) signalling. Mechanistically, GOLPH3 regulates cell size, enhances growth-factor-induced mTOR (also known as FRAP1) signalling in human cancer cells, and alters the response to an mTOR inhibitor in vivo. Thus, genomic and genetic, biological, functional and biochemical data in yeast and humans establishes GOLPH3 as a new oncogene that is commonly targeted for amplification in human cancer, and is capable of modulating the response to rapamycin, a cancer drug in clinical use.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Manning, B. D. & Cantley, L. C. AKT/PKB signaling: navigating downstream. Cell 129, 1261–1274 (2007)
Wullschleger, S., Loewith, R. & Hall, M. N. TOR signaling in growth and metabolism. Cell 124, 471–484 (2006)
Shima, H. et al. Disruption of the p70s6k/p85s6k gene reveals a small mouse phenotype and a new functional S6 kinase. EMBO J. 17, 6649–6659 (1998)
Montagne, J. et al. Drosophila S6 kinase: a regulator of cell size. Science 285, 2126–2129 (1999)
Oldham, S., Montagne, J., Radimerski, T., Thomas, G. & Hafen, E. Genetic and biochemical characterization of dTOR, the Drosophila homolog of the target of rapamycin. Genes Dev. 14, 2689–2694 (2000)
Zhang, H., Stallock, J. P., Ng, J. C., Reinhard, C. & Neufeld, T. P. Regulation of cellular growth by the Drosophila target of rapamycin dTOR. Genes Dev. 14, 2712–2724 (2000)
Fingar, D. C. & Blenis, J. Target of rapamycin (TOR): an integrator of nutrient and growth factor signals and coordinator of cell growth and cell cycle progression. Oncogene 23, 3151–3171 (2004)
Guertin, D. A. & Sabatini, D. M. Defining the role of mTOR in cancer. Cancer Cell 12, 9–22 (2007)
Yang, Q. & Guan, K. L. Expanding mTOR signaling. Cell Res. 17, 666–681 (2007)
Abraham, R. T. & Wiederrecht, G. J. Immunopharmacology of rapamycin. Annu. Rev. Immunol. 14, 483–510 (1996)
Sabatini, D. M. mTOR and cancer: insights into a complex relationship. Nature Rev. Cancer 6, 729–734 (2006)
Garraway, L. A. et al. Integrative genomic analyses identify MITF as a lineage survival oncogene amplified in malignant melanoma. Nature 436, 117–122 (2005)
Wu, C. C. et al. GMx33: a novel family of trans-Golgi proteins identified by proteomics. Traffic 1, 963–975 (2000)
Bell, A. W. et al. Proteomics characterization of abundant Golgi membrane proteins. J. Biol. Chem. 276, 5152–5165 (2001)
Snyder, C. M., Mardones, G. A., Ladinsky, M. S. & Howell, K. E. GMx33 associates with the trans-Golgi matrix in a dynamic manner and sorts within tubules exiting the Golgi. Mol. Biol. Cell 17, 511–524 (2006)
Bonifacino, J. S. & Hurley, J. H. Retromer. Curr. Opin. Cell Biol. 20, 427–436 (2008)
Xie, M. W. et al. Insights into TOR function and rapamycin response: chemical genomic profiling by using a high-density cell array method. Proc. Natl Acad. Sci. USA 102, 7215–7220 (2005)
Camp, R. L., Chung, G. G. & Rimm, D. L. Automated subcellular localization and quantification of protein expression in tissue microarrays. Nature Med. 8, 1323–1327 (2002)
Fingar, D. C., Salama, S., Tsou, C., Harlow, E. & Blenis, J. Mammalian cell size is controlled by mTOR and its downstream targets S6K1 and 4EBP1/eIF4E. Genes Dev. 16, 1472–1487 (2002)
Burnett, P. E., Barrow, R. K., Cohen, N. A., Snyder, S. H. & Sabatini, D. M. RAFT1 phosphorylation of the translational regulators p70 S6 kinase and 4E-BP1. Proc. Natl Acad. Sci. USA 95, 1432–1437 (1998)
Isotani, S. et al. Immunopurified mammalian target of rapamycin phosphorylates and activates p70 S6 kinase α in vitro . J. Biol. Chem. 274, 34493–34498 (1999)
Hresko, R. C. & Mueckler, M. mTOR·RICTOR is the Ser473 kinase for Akt/protein kinase B in 3T3–L1 adipocytes. J. Biol. Chem. 280, 40406–40416 (2005)
Sarbassov, D. D., Guertin, D. A., Ali, S. M. & Sabatini, D. M. Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science 307, 1098–1101 (2005)
Chiu, V. K. et al. Ras signalling on the endoplasmic reticulum and the Golgi. Nature Cell Biol. 4, 343–350 (2002)
Eaton, S. Retromer retrieves wntless. Dev. Cell 14, 4–6 (2008)
Korolchuk, V. I. et al. Drosophila Vps35 function is necessary for normal endocytic trafficking and actin cytoskeleton organisation. J. Cell Sci. 120, 4367–4376 (2007)
Schmitz, K. R. et al. Golgi localization of glycosyltransferases requires a Vps74p oligomer. Dev. Cell 14, 523–534 (2008)
Tu, L., Tai, W. C., Chen, L. & Banfield, D. K. Signal-mediated dynamic retention of glycosyltransferases in the Golgi. Science 321, 404–407 (2008)
Ohtsubo, K. & Marth, J. D. Glycosylation in cellular mechanisms of health and disease. Cell 126, 855–867 (2006)
Takahashi, M., Tsuda, T., Ikeda, Y., Honke, K. & Taniguchi, N. Role of N-glycans in growth factor signaling. Glycoconj. J. 20, 207–212 (2004)
Maser, R. S. et al. Chromosomally unstable mouse tumours have genomic alterations similar to diverse human cancers. Nature 447, 966–971 (2007)
Satyamoorthy, K. et al. Melanoma cell lines from different stages of progression and their biological and molecular analyses. Melanoma Res. 7 (suppl. 2). S35–S42 (1997)
Tonon, G. et al. High-resolution genomic profiles of human lung cancer. Proc. Natl Acad. Sci. USA 102, 9625–9630 (2005)
Acknowledgements
We thank R. DePinho for critical reading of the manuscript, and L. Cantley, as well as members of the Chin laboratory, for helpful discussion. We thank H. Ying for assistance with confocal microscopy. The rabbit polyclonal antibody against human GOLPH3 was kindly provided by J. J. Bergeron of McGill University. Mouse monoclonal antibody against human GOLPH3, C19, was generated at the Dana-Farber/Harvard Cancer Center Monoclonal Antibody Core Facility. K.L.S. is at present supported by a Postdoctoral Fellowship from the American Cancer Society (PF-07-039-01-CSM), and K.L.S. and O.K. were previously supported by a National Institutes of Health (NIH) Training Grant appointment in the Department of Dermatology at Brigham and Women’s Hospital (5-T32-AR07098-31). K.-K.W. was supported by a Program of Research Excellence (SPORE) grant (P50 CA090578) and NIH grants (R01 AG2400401; R01 CA122794). The AQUA immunofluorescence study was supported by a grant from the NIH to D.L.R. (RO-1 CA 114277). This work is primarily supported by grants from the NIH to L.C. (RO1 CA93947; P50 CA93683).
Author Contribution O.K. identified GOLPH3 as an oncogene target of 5p13; K.L.S. performed oncogene validation and mechanistic studies; E.I. and A.P. performed TMA FISH analysis; H.R.W. and D.E.F. performed the HMEL anchorage-independent growth assay; S.D. and J.W. assisted with immunofluorescence assays and provided technical support; M.W. performed the co-immunoprecipitation assays; V.A. and D.L.R. performed AQUA analysis; S.C. assisted with cell culture studies; A.S. aided phospholipid and biochemical analyses; M.-C.L. and K.-K.W. performed rapamycin xenograft assays; Y.X. performed computational analyses; T.F. and J.H. provided technical support; L.C. supervised the experiments and data interpretation; K.L.S. and L.C. wrote the manuscript.
Author information
Authors and Affiliations
Corresponding author
Supplementary information
Supplementary Information
This file contains Supplementary Tables 1-4 and Supplementary Figures S1-S5 with Legends. (PDF 428 kb)
Rights and permissions
About this article
Cite this article
Scott, K., Kabbarah, O., Liang, MC. et al. GOLPH3 modulates mTOR signalling and rapamycin sensitivity in cancer. Nature 459, 1085–1090 (2009). https://doi.org/10.1038/nature08109
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nature08109
This article is cited by
-
Golgi phosphoprotein 3 promotes angiogenesis and sorafenib resistance in hepatocellular carcinoma via upregulating exosomal miR-494-3p
Cancer Cell International (2022)
-
GOLPH3 protein controls organ growth by interacting with TOR signaling proteins in Drosophila
Cell Death & Disease (2022)
-
Golgi phosphoprotein 3 induces autophagy and epithelial–mesenchymal transition to promote metastasis in colon cancer
Cell Death Discovery (2022)
-
The understudied links of the retromer complex to age-related pathways
GeroScience (2022)
-
GOLPH3/CKAP4 promotes metastasis and tumorigenicity by enhancing the secretion of exosomal WNT3A in non-small-cell lung cancer
Cell Death & Disease (2021)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.