Photo of Stephen C. Blacklow,  MD, PhD

Stephen C. Blacklow, MD, PhD

Dana-Farber Cancer Institute

Dana-Farber Cancer Institute
Phone: (617) 525-4415
Fax: (617) 525-4414

Stephen C. Blacklow, MD, PhD

Dana-Farber Cancer Institute


  • Gustavus Adolphus Pfeiffer Professor of Biological Chemistry and Molecular Pharmacology, Biological Chemistry and Molecular Pharmacology, Harvard Medical School
  • Professor, Pathology, Brigham And Women's Hospital
  • Professor, Cancer Biology, Dana-Farber Cancer Institute
  • Chair, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School


Research Abstract


Keywords: lipoprotein receptor, protein structure, Notch, Notch1, nuclear magnetic resonance, protein dynamics.

The long-term goal of our research program is to elucidate the detailed molecular basis for specificity in protein-protein and receptor-ligand interactions, focusing on proteins implicated in human disease. Over the next several years, the laboratory will continue to explore structure-function relationships in proteins of the vascular and hematopoetic systems, with particular emphasis on lipoprotein and Notch receptors.

Lipoprotein receptors. The LDL receptor (LDLR) is the primary mechanism for uptake of cholesterol-carrying lipoprotein particles into cells, and serves as a prototype for a large family of cell surface receptors implicated in biological processes ranging from lipoprotein uptake to Wnt signal transduction. The basis for ligand recognition by the LDLR and related receptors remains poorly understood. We intend to elucidate at a detailed biochemical level how lipoprotein receptors bind to and release their ligands, and understand how other related receptors of this family transmit biological signals.

Mechanism of signal transduction by the human Notch1 receptor. Human Notch1 is a modular, single-pass transmembrane receptor that normally controls cellular differentiation in hematopoetic (and other) cells. The current working model for signaling by Notch receptors is that ligand binding triggers a cascade of proteolytic cleavages that release the intracellular portion of Notch from the membrane, allowing it to migrate to the nucleus where it activates transcription of Notch-responsive genes. The overarching goal of our studies is to elucidate the molecular logic of Notch signaling in different developmental, physiologic, and pathophysiologic contexts. To achieve this goal, we are combining biochemical, molecular, and structural approaches to gain insight into the mechanism of activation of Notch signaling. Our studies focus on three key steps in activation of Notch signals: how does the receptor recognize ligands, how is metalloprotease cleavage prevented prior to ligand binding, and how do the nuclear complexes assemble to activate transcription of target genes?


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  • Guo B, McMillan BJ, Blacklow SC. Structure and function of the Mind bomb E3 ligase in the context of Notch signal transduction. Curr Opin Struct Biol 2016; 41:38-45. PubMed
  • LaRochelle JR, Fodor M, Xu X, Durzynska I, Fan L, Stams T, Chan HM, LaMarche MJ, Chopra R, Wang P, Fortin PD, Acker MG, Blacklow SC. Structural and Functional Consequences of Three Cancer-Associated Mutations of the Oncogenic Phosphatase SHP2. Biochemistry 2016; 55:2269-77. PubMed
  • Uljon S, Xu X, Durzynska I, Stein S, Adelmant G, Marto JA, Pear WS, Blacklow SC. Structural Basis for Substrate Selectivity of the E3 Ligase COP1. Structure 2016. PubMed
  • Meyer PA, Socias S, Key J, Ransey E, Tjon EC, Buschiazzo A, Lei M, Botka C, Withrow J, Neau D, Rajashankar K, Anderson KS, Baxter RH, Blacklow SC, Boggon TJ, Bonvin AM, Borek D, Brett TJ, Caflisch A, Chang CI, Chazin WJ, Corbett KD, Cosgrove MS, Crosson S, Dhe-Paganon S, Di Cera E, Drennan CL, Eck MJ, Eichman BF, Fan QR, Ferré-D'Amaré AR, Christopher Fromme J, Garcia KC, Gaudet R, Gong P, Harrison SC, Heldwein EE, Jia Z, Keenan RJ, Kruse AC, Kvansakul M, McLellan JS, Modis Y, Nam Y, Otwinowski Z, Pai EF, Pereira PJ, Petosa C, Raman CS, Rapoport TA, Roll-Mecak A, Rosen MK, Rudenko G, Schlessinger J, Schwartz TU, Shamoo Y, Sondermann H, Tao YJ, Tolia NH, Tsodikov OV, Westover KD, Wu H, Foster I, Fraser JS, Maia FR, Gonen T, Kirchhausen T, Diederichs K, Crosas M, Sliz P. Data publication with the structural biology data grid supports live analysis. Nat Commun 2016; 7:10882. PubMed
  • Xu X, Choi SH, Hu T, Tiyanont K, Habets R, Groot AJ, Vooijs M, Aster JC, Chopra R, Fryer C, Blacklow SC. Insights into Autoregulation of Notch3 from Structural and Functional Studies of Its Negative Regulatory Region. Structure 2015; 23:1227-35. PubMed
  • Gordon WR, Zimmerman B, He L, Miles LJ, Huang J, Tiyanont K, McArthur DG, Aster JC, Perrimon N, Loparo JJ, Blacklow SC. Mechanical Allostery: Evidence for a Force Requirement in the Proteolytic Activation of Notch. Dev Cell 2015; 33:729-36. PubMed
  • Habets RA, Groot AJ, Yahyanejad S, Tiyanont K, Blacklow SC, Vooijs M. Human NOTCH2 Is Resistant to Ligand-independent Activation by Metalloprotease Adam17. J Biol Chem 2015; 290:14705-16. PubMed
  • McMillan BJ, Schnute B, Ohlenhard N, Zimmerman B, Miles L, Beglova N, Klein T, Blacklow SC. A tail of two sites: a bipartite mechanism for recognition of notch ligands by mind bomb E3 ligases. Mol Cell 2015; 57:912-24. PubMed
  • Yashiro-Ohtani Y, Wang H, Zang C, Arnett KL, Bailis W, Ho Y, Knoechel B, Lanauze C, Louis L, Forsyth KS, Chen S, Chung Y, Schug J, Blobel GA, Liebhaber SA, Bernstein BE, Blacklow SC, Liu XS, Aster JC, Pear WS. Long-range enhancer activity determines Myc sensitivity to Notch inhibitors in T cell leukemia. Proc Natl Acad Sci U S A 2014; 111:E4946-53. PubMed
  • McKeown MR, Shaw DL, Fu H, Liu S, Xu X, Marineau JJ, Huang Y, Zhang X, Buckley DL, Kadam A, Zhang Z, Blacklow SC, Qi J, Zhang W, Bradner JE. Biased multicomponent reactions to develop novel bromodomain inhibitors. J Med Chem 2014. PubMed
  • Wang H, Zang C, Taing L, Arnett KL, Wong YJ, Pear WS, Blacklow SC, Liu XS, Aster JC. NOTCH1-RBPJ complexes drive target gene expression through dynamic interactions with superenhancers. Proc Natl Acad Sci U S A 2014; 111:705-10. PubMed
  • Blacklow SC. Refining a Jagged edge. Structure 2013; 21:2100-1. PubMed
  • Tiyanont K, Wales TE, Siebel CW, Engen JR, Blacklow SC. Insights into Notch3 activation and inhibition mediated by antibodies directed against its negative regulatory region. J Mol Biol 2013; 425:3192-204. PubMed
  • Andrawes MB, Xu X, Liu H, Ficarro SB, Marto JA, Aster JC, Blacklow SC. Intrinsic selectivity of Notch 1 for Delta-like 4 over Delta-like 1. J Biol Chem 2013; 288:25477-89. PubMed
  • Roti G, Carlton A, Ross KN, Markstein M, Pajcini K, Su AH, Perrimon N, Pear WS, Kung AL, Blacklow SC, Aster JC, Stegmaier K. Complementary genomic screens identify SERCA as a therapeutic target in NOTCH1 mutated cancer. Cancer Cell 2013; 23:390-405. PubMed
  • Calderwood MA, Lee S, Holthaus AM, Blacklow SC, Kieff E, Johannsen E. Epstein-Barr virus nuclear protein 3C binds to the N-terminal (NTD) and beta trefoil domains (BTD) of RBP/CSL; only the NTD interaction is essential for lymphoblastoid cell growth. Virology 2011. PubMed
  • Aster JC, Blacklow SC, Pear WS. Notch signalling in T-cell lymphoblastic leukaemia/lymphoma and other haematological malignancies. J Pathol 2010; 223:262-73. PubMed
  • Ashworth TD, Pear WS, Chiang MY, Blacklow SC, Mastio J, Xu L, Kelliher M, Kastner P, Chan S, Aster JC. Deletion-based mechanisms of Notch1 activation in T-ALL: key roles for RAG recombinase and a conserved internal translational start site in Notch1. Blood 2010; 116:5455-64. PubMed
  • Keeshan K, Bailis W, Dedhia PH, Vega ME, Shestova O, Xu L, Toscano K, Uljon SN, Blacklow SC, Pear WS. Transformation by Tribbles homologue 2 (Trib2) requires both the Trib2 kinase domain and COP1 binding. Blood 2010; 116:4948-57. PubMed
  • Arnett KL, Hass M, McArthur DG, Ilagan MX, Aster JC, Kopan R, Blacklow SC. Structural and mechanistic insights into cooperative assembly of dimeric Notch transcription complexes. Nat Struct Mol Biol 2010; 17:1312-7. PubMed
  • Liu H, Chi AW, Arnett KL, Chiang MY, Xu L, Shestova O, Wang H, Li YM, Bhandoola A, Aster JC, Blacklow SC, Pear WS. Notch dimerization is required for leukemogenesis and T-cell development. Genes Dev 2010; 24:2395-407. PubMed
  • Dedhia PH, Keeshan K, Uljon S, Xu L, Vega ME, Shestova O, Zaks-Zilberman M, Romany C, Blacklow SC, Pear WS. Differential ability of Tribbles family members to promote degradation of C/EBP{alpha} and induce acute myelogenous leukemia. Blood 2010; 116:1321-8. PubMed
  • Kovall RA, Blacklow SC. Mechanistic insights into Notch receptor signaling from structural and biochemical studies. Curr Top Dev Biol 2010; 92:31-71. PubMed
  • Aste-Amézaga M, Zhang N, Lineberger JE, Arnold BA, Toner TJ, Gu M, Huang L, Vitelli S, Vo KT, Haytko P, Zhao JZ, Baleydier F, L'Heureux S, Wang H, Gordon WR, Thoryk E, Andrawes MB, Tiyanont K, Stegmaier K, Roti G, Ross KN, Franlin LL, Wang H, Wang F, Chastain M, Bett AJ, Audoly LP, Aster JC, Blacklow SC, Huber HE. Characterization of Notch1 antibodies that inhibit signaling of both normal and mutated Notch1 receptors. PLoS ONE 2010; 5:e9094. PubMed
  • Del Bianco C, Vedenko A, Choi SH, Berger MF, Shokri L, Bulyk ML, Blacklow SC. Notch and MAML-1 complexation do not detectably alter the dna binding specificity of the transcription factor CSL. PLoS ONE 2010; 5:e15034. PubMed
  • Moellering RE, Cornejo M, Davis TN, Del Bianco C, Aster JC, Blacklow SC, Kung AL, Gilliland DG, Verdine GL, Bradner JE. Direct inhibition of the NOTCH transcription factor complex. Nature 2009; 462:182-8. PubMed
  • Yashiro-Ohtani Y, He Y, Ohtani T, Jones ME, Shestova O, Xu L, Fang TC, Chiang MY, Intlekofer AM, Blacklow SC, Zhuang Y, Pear WS. Pre-TCR signaling inactivates Notch1 transcription by antagonizing E2A. Genes Dev 2009; 23:1665-76. PubMed
  • Gordon WR,Roy M,Vardar-Ulu D,Garfinkel M,Mansour MR,Aster JC,Blacklow SC. Structure of the Notch1-negative regulatory region: implications for normal activation and pathogenic signaling in T-ALL. Blood 2008; 113:4381-90. PubMed
  • Gordon WR, Vardar-Ulu D, L'Heureux S, Ashworth T, Malecki MJ, Sanchez-Irizarry C, McArthur DG, Histen G, Mitchell JL, Aster JC, Blacklow SC. Effects of S1 cleavage on the structure, surface export, and signaling activity of human Notch1 and Notch2. PLoS ONE 2009; 4:e6613. PubMed
  • Gordon WR,Arnett KL,Blacklow SC. The molecular logic of Notch signaling--a structural and biochemical perspective. J Cell Sci 2008; 121:3109-19. PubMed
  • Li K, Li Y, Wu W, Gordon WR, Chang DW, Lu M, Scoggin S, Fu T, Vien L, Histen G, Zheng J, Martin-Hollister R, Duensing T, Singh S, Blacklow SC, Yao Z, Aster JC, Zhou BB. Modulation of Notch signaling by antibodies specific for the extracellular negative regulatory region of NOTCH3. J Biol Chem 2008; 283:8046-54. PubMed
  • Estrada K, Fisher C, Blacklow SC. Unfolding of the RAP-D3 helical bundle facilitates dissociation of RAP-receptor complexes. Biochemistry 2008; 47:1532-9. PubMed
  • Del Bianco C, Aster JC, Blacklow SC. Mutational and energetic studies of Notch 1 transcription complexes. J Mol Biol 2007; 376:131-40. PubMed
  • Aster JC, Pear WS, Blacklow SC. Notch signaling in leukemia. Annu Rev Pathol 2007; 3:587-613. PubMed
  • Blacklow SC. Versatility in ligand recognition by LDL receptor family proteins: advances and frontiers. Curr Opin Struct Biol 2007; 17:419-26. PubMed
  • Fang TC, Yashiro-Ohtani Y, Del Bianco C, Knoblock DM, Blacklow SC, Pear WS. Notch directly regulates Gata3 expression during T helper 2 cell differentiation. Immunity 2007; 27:100-10. PubMed
  • Koduri V, Blacklow SC. Requirement for natively unstructured regions of mesoderm development candidate 2 in promoting low-density lipoprotein receptor-related protein 6 maturation. Biochemistry 2007; 46:6570-7. PubMed
  • Gordon WR, Vardar-Ulu D, Histen G, Sanchez-Irizarry C, Aster JC, Blacklow SC. Corrigendum: Structural basis for autoinhibition of Notch. Nat Struct Mol Biol 2007; 14:455. PubMed
  • Gordon WR, Vardar-Ulu D, Histen G, Sanchez-Irizarry C, Aster JC, Blacklow SC. Structural basis for autoinhibition of Notch. Nat Struct Mol Biol 2007; 14:295-300. PubMed
  • Nam Y, Sliz P, Pear WS, Aster JC, Blacklow SC. Cooperative assembly of higher-order Notch complexes functions as a switch to induce transcription. Proc Natl Acad Sci U S A 2007; 104:2103-8. PubMed
  • Weng AP, Millholland JM, Yashiro-Ohtani Y, Arcangeli ML, Lau A, Wai C, Del Bianco C, Rodriguez CG, Sai H, Tobias J, Li Y, Wolfe MS, Shachaf C, Felsher D, Blacklow SC, Pear WS, Aster. c-Myc is an important direct target of Notch1 in T-cell acute lymphoblastic leukemia/lymphoma. Genes Dev 2006; 20:2096-109. PubMed
  • Chiang MY, Xu ML, Histen G, Shestova O, Roy M, Nam Y, Blacklow SC, Sacks DB, Pear WS, Aster JC. Identification of a Conserved Negative Regulatory Sequence That Influences the Leukemogenic Activity of NOTCH1. Mol Cell Biol 2006; 26:6261-71. PubMed
  • Malecki MJ, Sanchez-Irizarry C, Mitchell JL, Histen G, Xu ML, Aster JC, Blacklow SC. Leukemia-associated mutations within the NOTCH1 heterodimerization domain fall into at least two distinct mechanistic classes. Mol Cell Biol 2006; 26:4642-51. PubMed
  • Lee D, Walsh JD, Mikhailenko I, Yu P, Migliorini M, Wu Y, Krueger S, Curtis JE, Harris B, Lockett S, Blacklow SC, Strickland DK, Wang YX. RAP uses a histidine switch to regulate its interaction with LRP in the ER and Golgi. Mol Cell 2006; 22:423-30. PubMed
  • Fisher C, Beglova N, Blacklow SC. Structure of an LDLR-RAP complex reveals a general mode for ligand recognition by lipoprotein receptors. Mol Cell 2006; 22:277-83. PubMed
  • Nam Y, Sliz P, Song L, Aster JC, Blacklow SC. Structural basis for cooperativity in recruitment of MAML coactivators to Notch transcription complexes. Cell 2006; 124:973-83. PubMed
  • Moellering RE, Cornejo M, Davis TN, Del Bianco C, Aster JC, Blacklow SC, Kung AL, Gilliland DG, Verdine GL, Bradner J. Direct inhibition of the Notch transcription factor complex Nature 2009.