Abstract
Activating mutations in genes encoding G protein α (Gα) subunits occur in 4–5% of all human cancers1, but oncogenic alterations in Gβ subunits have not been defined. Here we demonstrate that recurrent mutations in the Gβ proteins GNB1 and GNB2 confer cytokine-independent growth and activate canonical G protein signaling. Multiple mutations in GNB1 affect the protein interface that binds Gα subunits as well as downstream effectors and disrupt Gα interactions with the Gβγ dimer. Different mutations in Gβ proteins clustered partly on the basis of lineage; for example, all 11 GNB1 K57 mutations were in myeloid neoplasms, and seven of eight GNB1 I80 mutations were in B cell neoplasms. Expression of patient-derived GNB1 variants in Cdkn2a-deficient mouse bone marrow followed by transplantation resulted in either myeloid or B cell malignancies. In vivo treatment with the dual PI3K-mTOR inhibitor BEZ235 suppressed GNB1-induced signaling and markedly increased survival. In several human tumors, mutations in the gene encoding GNB1 co-occurred with oncogenic kinase alterations, including the BCR-ABL fusion protein, the V617F substitution in JAK2 and the V600K substitution in BRAF. Coexpression of patient-derived GNB1 variants with these mutant kinases resulted in inhibitor resistance in each context. Thus, GNB1 and GNB2 alterations confer transformed and resistance phenotypes across a range of human tumors and may be targetable with inhibitors of G protein signaling.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 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
O'Hayre, M. et al. The emerging mutational landscape of G proteins and G-protein-coupled receptors in cancer. Nat. Rev. Cancer 13, 412–424 (2013).
Garraway, L.A. & Lander, E.S. Lessons from the cancer genome. Cell 153, 17–37 (2013).
Lawrence, M.S. et al. Discovery and saturation analysis of cancer genes across 21 tumour types. Nature 505, 495–501 (2014).
Iyer, G. et al. Genome sequencing identifies a basis for everolimus sensitivity. Science 338, 221 (2012).
Yoda, A. et al. Functional screening identifies CRLF2 in precursor B-cell acute lymphoblastic leukemia. Proc. Natl. Acad. Sci. USA 107, 252–257 (2010).
Shindoh, N. et al. Next-generation cDNA screening for oncogene and resistance phenotypes. PLoS ONE 7, e49201 (2012).
Lucioni, M. et al. Twenty-one cases of blastic plasmacytoid dendritic cell neoplasm: focus on biallelic locus 9p21.3 deletion. Blood 118, 4591–4594 (2011).
Menezes, J. et al. Exome sequencing reveals novel and recurrent mutations with clinical impact in blastic plasmacytoid dendritic cell neoplasm. Leukemia 28, 823–829 (2014).
Oldham, W.M. & Hamm, H.E. Heterotrimeric G protein activation by G-protein-coupled receptors. Nat. Rev. Mol. Cell Biol. 9, 60–71 (2008).
Radhika, V. & Dhanasekaran, N. Transforming G proteins. Oncogene 20, 1607–1614 (2001).
Van Raamsdonk, C.D. et al. Frequent somatic mutations of GNAQ in uveal melanoma and blue naevi. Nature 457, 599–602 (2009).
Vallar, L., Spada, A. & Giannattasio, G. Altered Gs and adenylate cyclase activity in human GH-secreting pituitary adenomas. Nature 330, 566–568 (1987).
Walter, M.J. et al. Clonal diversity of recurrently mutated genes in myelodysplastic syndromes. Leukemia 27, 1275–1282 (2013).
Haferlach, T. et al. Landscape of genetic lesions in 944 patients with myelodysplastic syndromes. Leukemia 28, 241–247 (2014).
Ford, C.E. et al. Molecular basis for interactions of G protein βγ subunits with effectors. Science 280, 1271–1274 (1998).
Wall, M.A. et al. The structure of the G protein heterotrimer Giα1β1γ2 . Cell 83, 1047–1058 (1995).
Adelmant, G. et al. DNA ends alter the molecular composition and localization of Ku multicomponent complexes. Mol. Cell Proteomics 11, 411–421 (2012).
Willardson, B.M. & Tracy, C.M. Chaperone-mediated assembly of G protein complexes. Subcell. Biochem. 63, 131–153 (2012).
Hakak, Y. et al. The role of the GPR91 ligand succinate in hematopoiesis. J. Leukoc. Biol. 85, 837–843 (2009).
Gupta, S.K. et al. Analysis of the fibroblast transformation potential of GTPase-deficient gip2 oncogenes. Mol. Cell. Biol. 12, 190–197 (1992).
Stephens, L. et al. A novel phosphoinositide 3 kinase activity in myeloid-derived cells is activated by G protein βγ subunits. Cell 77, 83–93 (1994).
Crespo, P., Xu, N., Simonds, W.F. & Gutkind, J.S. Ras-dependent activation of MAP kinase pathway mediated by G-protein βγ subunits. Nature 369, 418–420 (1994).
Camps, M. et al. Isozyme-selective stimulation of phospholipase C-β2 by G protein βγ subunits. Nature 360, 684–686 (1992).
Subramanian, A. et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl. Acad. Sci. USA 102, 15545–15550 (2005).
Krause, D.S., Lazarides, K., von Andrian, U.H. & Van Etten, R.A. Requirement for CD44 in homing and engraftment of BCR-ABL-expressing leukemic stem cells. Nat. Med. 12, 1175–1180 (2006).
Akagi, T. et al. Frequent genomic abnormalities in acute myeloid leukemia/myelodysplastic syndrome with normal karyotype. Haematologica 94, 213–223 (2009).
Sherborne, A.L. et al. Variation in CDKN2A at 9p21.3 influences childhood acute lymphoblastic leukemia risk. Nat. Genet. 42, 492–494 (2010).
Tyner, J.W. et al. Kinase pathway dependence in primary human leukemias determined by rapid inhibitor screening. Cancer Res. 73, 285–296 (2013).
Johannessen, C.M. et al. A melanocyte lineage program confers resistance to MAP kinase pathway inhibition. Nature 504, 138–142 (2013).
Chapman, P.B. et al. Improved survival with vemurafenib in melanoma with BRAF V600E mutation. N. Engl. J. Med. 364, 2507–2516 (2011).
van Bodegom, D. et al. Differences in signaling through the B-cell leukemia oncoprotein CRLF2 in response to TSLP and through mutant JAK2. Blood 120, 2853–2863 (2012).
Ficarro, S.B. et al. Magnetic bead processor for rapid evaluation and optimization of parameters for phosphopeptide enrichment. Anal. Chem. 81, 4566–4575 (2009).
Ficarro, S.B. et al. Online nanoflow multidimensional fractionation for high-efficiency phosphopeptide analysis. Mol. Cell Proteomics 10, O111.011064 (2011).
Askenazi, M., Parikh, J.R. & Marto, J.A. mzAPI: a new strategy for efficiently sharing mass spectrometry data. Nat. Methods 6, 240–241 (2009).
Parikh, J.R. et al. multiplierz: an extensible API based desktop environment for proteomics data analysis. BMC Bioinformatics 10, 364 (2009).
Askenazi, M., Marto, J.A. & Linial, M. The complete peptide dictionary—a meta-proteomics resource. Proteomics 10, 4306–4310 (2010).
Chapuy, B. et al. Discovery and characterization of super-enhancer-associated dependencies in diffuse large B cell lymphoma. Cancer Cell 24, 777–790 (2013).
Weigert, O. et al. Genetic resistance to JAK2 enzymatic inhibitors is overcome by HSP90 inhibition. J. Exp. Med. 209, 259–273 (2012).
Elpek, K.G. et al. Lymphoid organ-resident dendritic cells exhibit unique transcriptional fingerprints based on subset and site. PLoS ONE 6, e23921 (2011).
Acknowledgements
We thank T. Radimerski, T. Haferlach, L. Garraway, M. Walter and N. Wagle for contributing to the manuscript. We thank A. Hawkins and the Brigham and Women's Hospital Cytogenomics Core Facility for karyotyping. This research was supported by the Aplastic Anemia and MDS International Foundation (A.Y.), the Edwards P. Evans Foundation (A.Y.), the Max-Eder Program of the Deutsche Krebshilfe (#110659 to O.W.), the Leukemia and Lymphoma Society (J.W.T.), US National Institutes of Health NCI (1R01CA183947 and 5R00CA151457 to J.W.T.), the V Foundation for Cancer Research (J.W.T.), Gabrielle's Angel Foundation for Cancer Research (J.W.T.), the Claudia Adams Barr Fund (A.Y. and D.M.W.), an American Cancer Society Research Scholar Grant (D.M.W.), a Stand Up To Cancer Innovative Research Grant (D.M.W.), and an American Society of Hematology Scholar Award (A.A.L.).
Author information
Authors and Affiliations
Contributions
A.Y., G.A., J.T., B.C., N.S., Y.Y., O.W., N.K., S.-C.W., S.S.K., H.L., T.T., A.L.C., K.G.E., J.C., N.J.-S. and A.A.L. designed and performed experiments. A.Y., G.A., J.T., B.C., K.G., S.J.T., S.J.R., J.W.T., J.A.M., D.M.W. and A.A.L. analyzed data. J.G., M.W.D., H.M., J.P.M., S.J. and B.L.E. provided essential reagents. A.Y., D.M.W. and A.A.L. wrote the paper.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–7 (PDF 78215 kb)
Supplementary Table 1
List of mutations in GNB1 and GNB2 (XLSX 16 kb)
Supplementary Table 2
Detected proteins from a band unique to the FLAG-GNB1 immuoprecipitation (XLSX 11 kb)
Supplementary Table 3
List of proteins from tandem affinity purification and mass spectrometry analysis (XLSX 28 kb)
Supplementary Table 4
List of proteins detected by phosphoproteomics analysis (XLSX 2508 kb)
Supplementary Table 5
List of phospho-sites with more than 1.5 times enrichment in TF1-GNB1 K89E cells compared to TF1 cells infected with an empty vector (XLSX 53 kb)
Supplementary Table 6
Drug screening of GNB1 K89E cells (XLSX 13 kb)
Supplementary Table 7
Assembled G protein downstream signaling pathway gene sets from MSigDB queried by GSEA in Figure 3 (XLSX 599 kb)
Source data
Rights and permissions
About this article
Cite this article
Yoda, A., Adelmant, G., Tamburini, J. et al. Mutations in G protein β subunits promote transformation and kinase inhibitor resistance. Nat Med 21, 71–75 (2015). https://doi.org/10.1038/nm.3751
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nm.3751
This article is cited by
-
Genetics and epidemiology of mutational barcode-defined clonal hematopoiesis
Nature Genetics (2023)
-
PAX8-AS1 knockdown facilitates cell growth and inactivates autophagy in osteoblasts via the miR-1252-5p/GNB1 axis in osteoporosis
Experimental & Molecular Medicine (2021)
-
Tumor-induced neurogenesis and immune evasion as targets of innovative anti-cancer therapies
Signal Transduction and Targeted Therapy (2020)
-
Molecular pathogenesis of disease progression in MLL-rearranged AML
Leukemia (2019)
-
Transcriptome 3′end organization by PCF11 links alternative polyadenylation to formation and neuronal differentiation of neuroblastoma
Nature Communications (2018)