M. Ben Major, Ph.D.

MAJOR_1

ben_major@med.unc.edu

Major Lab Web Page

Lab Personnel

  • Priscila Siesser, PhD
  • Matt Walker, PhD
  • Feng Yan, PhD
  • Erica Cloer
  • Dennis Goldfarb
  • Kathleen Mulvaney

Assistant Professor

  • B.S., Michigan State University, 1997
  • Ph.D., Huntsman Cancer Institute, 2004
  • Postdoc, University of Washington, 2004-2009
  • Joined the Department in 2009

Research Interests

Although initiated by genetic mutation, the unchecked proliferation, aberrant differentiation, and altered motility of cancer cells depends upon the integrity and activation state of specific signal transduction pathways. Our laboratory is interested in understanding how alterations in these signaling pathways contribute to human cancer, and whether exploitation of that understanding can aid in the development of new diagnostics, prognostics and therapeutic intervention strategies. To this end, we employ a global “systems level” integrative discovery platform, one that has as a foundation mass spectrometry-based proteomic interaction networks. More specifically, through LC/MS/MS, we define the physical interaction network for a signaling pathway of oncogenic interest. By small molecule and functional genomic screening, we then annotate the human genome for functional contribution to the pathway of interest. Integration of these data with cancer-associated mutation data and cancer-associated gene expression data yields a powerful tool for oncogenic discovery—a cancer annotated physical/functional map for a specific signaling pathway of interest. The models and hypotheses produced though integrative screening are challenged through mechanistic studies employing cultured human cancer cells, zebrafish, mice and in vitro biochemical systems. With this general approach, we are currently pursuing the following projects:

Wnt/β-catenin Signal Transduction. Of the relatively small number of signaling pathways that function as master regulators of development, adult tissue homeostasis and cancer, the b-catenin dependent Wnt pathway (Wnt/b-catenin) figures prominently; it regulates the growth and fate of neoplastic cells in tissues of diverse origin, notably the colon, kidney, breast and skin. Integrative molecular analysis of Wnt/b-catenin signaling in kidney and colon cancer models yielded numerous discoveries of mechanistic and clinical importance. Work in my lab is now focusing on an integrative analysis of Wnt signaling in B-cell lymphoma and lung cancer. For both cancer models, independent functional genomic screens have been completed and have identified numerous novel regulators, some of which are mutated in human cancer. In addition, we are looking more closely at a family of proteins I discovered to regulate Wnt signaling in Wilms tumor. Specifically, we are working to understand the mechanism(s) by which members of the WTX gene family regulate Wnt signaling and oncogenesis.

Keap1/Nrf2 Signal Transduction. Keap1 is an E3 ubiquitin ligase important for cellular defense against genotoxic stress, and in that context contributes fundamentally to aging and a myriad of human cancers, most notably lung cancer. Keap1 functions by ubiquitinating the Nrf2 transcription factor, which ultimately results in the proteosomal degradation of Nrf2. In an effort to better understand Keap1 in an oncogenic contect, I completed quantitative proteomic analysis of the Keap1 protein complex as well as functional genomic screen of the Keap1/Nrf2 pathway. The resulting integrative map has identified numerous novel proteins which both physically associate with keap1 as well as functional regulate Keap1/Nrf2 signaling. Ongoing work in the lab is focused on understanding the mechanisms by which these proteins control Keap1 function as well as uncovering new cancer connections.

E3 Ubiquitin Ligase Substrate Identification. E3 ubiquitin ligase complexes provide specificity and catalysis for the transfer of ubiquitin to target proteins, a post-translational modification that results in proteosome-mediated degradation, altered subcellular localization or changes in protein interaction. As such, E3 ubiquitin ligases regulate every facet of cell biology, and importantly, are frequently perturbed in disease states such as cancer. While expanding my protein-protein interaction network for the Wnt/b-catenin pathway, I performed tandem affinity purification and mass spectrometry on bTrCP, the E3 ubiquitin ligase responsible for b-catenin degration. As the E3 complex is catalytic in action, many of the known bTrCP substrates were not identified by LC/MS/MS. As such I designed and implemented a novel approach which stabilizes the E3-substrate interaction. Using this strategy, I have identified novel substrates for both the bTrCp and Keap1 E3 ubiquitin ligases. Work in my laboratory is exploiting this system to identify substrates for uncharacterized E3 ubiquitin ligases, specifically those with established connections to human disease.

Awards and Honors

2010:  Sidney Kimmel Scholar Award

2010:  NIH Directors New Innovator Award

Clinical Interests

  • non-small cell lung cancer
  • B-cell lymphoma

Selected Publications

PubMed 1

  • Bridgid E. Hast, Erica W. Cloer, Dennis Goldfarb, Heng Li, Priscila F. Siesser, Feng Yan, Vonn Walter, Ning Zheng, D. Neil Hayes and Michael B. Major. Cancer-derived mutations in KEAP1 impair NRF2 degradation but not ubiquitination. Cancer Research, 2013, Dec. 9, 2013;  doi:10.1158/0008-5472.CAN-13-1655
  • Bridgid E. Hast, Dennis Goldfarb, Kathleen M. Mulvaney, Michael A. Hast, Priscila F. Siesser, Feng Yan, D. Neil Hayes and Michael B. Major. Proteomic analysis of ubiquitin ligase KEAP1 reveals associated proteins that inhibit NRF2 ubiquitination. Cancer Research, Apr 1st, 2013;73(7):2199-210. 
  • Priscila F. Siesser, Marta Motolese, Matthew P. Walker, Dennis Goldfarb, Kelly Gewain, Feng Yan, Rima M. Kulikauskas, Andy J. Chien, Linda Wordeman and Michael B. Major. FAM123A Binds Microtubules and Inhibits the Guanine Nucleotide Exchange Factor ARHGEF2 to Decrease Actomyosin Contractility. Science Signaling. 2012 Sep 4; 240(5):ra64. PMID: 22949735
  • Shelly Sorrells, Seth Carbonneau, Erik Harrington, Aye T. Chen, Bridgid Hast, Brett Milash, Ujwal Pyati, Michael B. Major,Yi Zhou, Rodney A. Stewart, Leonard I. Zon, A. Thomas Look, and Cicely Jette. Ccdc94 Functions in the Prp19 Complex to Protect Cells from Radiation-Induced Apoptosis by Inhibiting the Transcription of p53. PLoS Genet. 2012 Aug 8; 8(8):e1002922. PMCID: PMC3431329
  • Duncan, J.S., Whittle, M.C., Nakamura, K., Abell, A.N., Midland, A.A., Zawistowski, J.S., Johnson, N.L., Granger, D.A., Vincent Jordan, N.,  Darr, D.B., Usary, J., Kuan, P.F., Smalley, D.M., Major, B., He, X., Hoadley, K., Zhou, B. Sharpless, N.E., Perou, C.M., Kim, W.Y.,  Gomez, S.H., Chen, X., Jin, J., Frye, S.V., Earp, H.S., Graves, L.M., Johnson, G.L. (2012) Dynamic Reprogramming of the Kinome In Response to Targeted MEK Inhibition In Triple Negative Breast Cancer. Cell. 2012 Apr 13; 149(2):307-21.  PMCID:PMC3328787
  • Camp ND, James RG, Dawson DW, Yan F, Davison JM, Houck SA, Tang X, Zheng N, Major MB*, Moon RT*.  Wilms tumor gene on the X chromosome (WTX) inhibits the degradation of NRF2 through comptetitive binding to KEAP1. Journal of Biological Chemistry. 2012 Jan 3. PMCID: PMC3307315
  • Chung N, Marine S, Smith EA, Liehr R, Smith ST, Locco L, Hudak E, Kreamer A, Rush A, Roberts B, Major MB, Moon RT, Arthur W, Cleary M, Strulovici B, Ferrer M.  A 1,536-well ultra-high-throughput siRNA screen to identify regulators of the Wnt/beta-catenin pathway. Assay Drug Dev Tech., Jun 2010:8(2):286-94. PMID:20578927
  • Jason D. Berndt, Travis L. Biechele, Randall T. Moon, and Michael B. Major. Integrative Analysis of Genome-Wide RNA Interference Screens. Science Signaling. 2009 12 May Vol. 2, Issue 70, p. pt4. PMID: 19436058
  • Richard G. James, Travis L. Biechele, William H. Conrad, Nathan D. Camp, Daniel M. Fass, Michael B. Major, Karen Sommer, XianHua Yi, Brian S. Roberts, Michelle A. Cleary, William T. Arthur, Michael MacCoss, David J. Rawlings, Stephen J. Haggerty and Randall T. Moon. Bruton’s Tyrosine Kinase Binds CDC73 and Negatively Regulates Wnt/b-catenin signaling. Science Signaling., 26 May 2009 Vol. 2, Issue 72, p. ra25. PMID: 19471023
  • Michael B. Major and Randall T. Moon. Risk Assessment in the Age of “Omics” Sci. Signal., 26 May 2009 Vol. 2, Issue 72, p. eg7
  • Richard G. James, Travis L. Biechele, William H. Conrad, Nathan D. Camp, Daniel M. Fass, Michael B. Major, Karen Sommer, XianHua Yi, Brian S. Roberts, Michelle A. Cleary, William T. Arthur, Michael MacCoss, David J. Rawlings, Stephen J. Haggerty and Randall T. Moon. Bruton’s Tyrosine Kinase Binds CDC73 and Negatively Regulates Wnt/b-catenin signaling. Sci. Signal., 26 May 2009 Vol. 2, Issue 72, p. ra25
  • Jason D. Berndt, Travis L. Biechele, Randall T. Moon, and Michael B. Major. Integrative Analysis of Genome-Wide RNA Interference Screens. Sci. Signal., 12 May 2009 Vol. 2, Issue 70, p. pt4
  • Jason D. Berndt, Randall T. Moon and Michael B. Major. b-Catenin gets Jaded, and it is VHL’s fault. Trends in Biological Science. 2009 Mar Vol. 34, p.101-104
  • Andy J. Chien, Erin C. Moore, Anke S. Lonsdorf, Rima M. Kulikauskas, Bonnie Gould Rothberg, Aaron J. Berger, Michael B. Major, Sam T. Hwang, David L. Rimm, and Randall T. Moon. Activated Wnt/b-catenin signaling in melanoma is associated with decreased proliferation in patient tumors and a murine melanoma model. Proceedings of the National Academy of Sciences. 2009 Jan 27;106(4):1193-1198.
  • Michael B. Major, Brian S. Roberts, Jason D. Berndt, Shane Marine, Jamie Anastas, Namjin Chung, Marc Ferrer, XianHua Yi, Cristi L. Stoick-Cooper, Priska D. von Haller, Lorna Kategaya, Andy Chien, Stephane Angers, Michael MacCoss, Michele A. Cleary, William T. Arthur and Randall T. Moon. An Integrative Molecular Screening Approach Identifies Regulators of Wnt/-catenin Signal Transduction. Science Signaling. 2008 Nov 11;1(45)
  • De Ferrari GV, Papassotiropoulos A, Biechele T, De-Vrieze FW, Avila ME, Major MB, Myers A, Sáez K, Henríquez JP, Zhao A, Wollmer MA, Nitsch RM, Hock C, Morris CM, Hardy J, Moon RT. Common genetic variation within the Low-Density Lipoprotein Receptor-Related Protein 6 and late-onset Alzheimer's disease. Proceedings of the National Academy of Sciences. 2007 May 29;104(22):9434-9439.
  • Michael B. Major, Nathan D. Camp, Jason D. Berndt, XianHua Yi, Seth J. Goldenberg, Charlotte Hubbert, Travis L. Biechele, Anne-Claude Gingras, Ning Zheng, Michael J. MacCoss, Stephane Angers, Randall T. Moon. Wilms Tumor Suppressor WTX Negatively Regulates Wnt/b-catenin Signaling. Science. 2007. May 18;316(5827):1043-1046.
  • Qisheng Zhang, Michael B. Major, Shinichi Takanashi, Nathan D. Camp, Naoyuki Nishiya, Eric C. Peters, Mark H. Ginsberg, Peter G. Schultz , Randall T. Moon, and Sheng Ding. Small-molecule synergist of the Wnt/b-catenin signaling pathway. Proceedings of the National Academy of Sciences. 2007 May 1;104(18):7444-7448
  • Cristi L. Stoick-Cooper, Gilbert Weidinger, Kimberly J. Riehle, Charlotte Hubbert, Michael B. Major, Nelson Fausto and Randall T. Moon. Distinct Wnt signaling pathways have opposing roles in appendage regeneration. Development. 2007. 134:479-489
  • Michael B. Major and David A. Jones. Identification of a Gadd45b 3-prime enhancer that mediates SMAD3 and SMAD4 dependent transcriptional induction by TGFb. Journal of Biological Chemistry. 2004 Feb 13;279(7):5278-5287.
  • Susanne Kloeker, Michael B. Major, David A. Calderwood, Mark H. Ginsberg, David A. Jones, and Mary C. Beckerle. The Kindler syndrome protein is regulated by TGFb and involved in integrin-mediated adhesion. Journal of Biological Chemistry. 2004 Feb 20;279(8):6824-33.