Stem Cell Center, Yale: Tissue Specific Stem Cells
Protein Tyrosine Phosphatases in Growth and Development.
The net cellular level of tyrosine phosphorylation is regulated by the intrinsic and opposing activities of protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs).
Protein tyrosine phosphorylation mediates numerous fundamental physiological events such as mitogenesis, differentiation, cell movement and apoptosis. Our laboratory is interested in how PTPs participate in the regulation of these cellular processes. In order to decipher how PTPs regulate mammalian cell signaling we use a broad range of approaches from molecular biology to mouse genetic strategies.
SH2 domain-containing PTPs - SHP-2.
SHP-2 is a ubiquitously expressed PTP containing tandem SH2 domains. We have been interested in understanding the role of SHP-2 in skeletal muscle growth and differentiation. We have shown that SHP-2 is critical for growth factor signaling in cultured muscle cells. The activity of SHP-2 also is important for cultured muscle cells to differentiate into multinucleated myotubes. Our research efforts are now focused on understanding the developmental and post-developmental roles of SHP-2 in skeletal muscle using mouse models.
In virtually all cases, the catalytic activity of SHP-2 is required for normal cellular function. However, the substrates that are dephosphorylated by SHP-2 are largely unknown. To identify SHP-2 substrates, we are using proteomic approaches in conjunction with mutants of SHP-2 that selectively bind to its substrate.
Mitogen-activated protein kinase (MAPK) phosphatases - MKP-1.
The activation of the MAPKs is critical for the control of a multitude of biological processes. We are interested in how the MAPKs are regulated by a family of PTPs known as MAPK phosphatases (MKPs) that inactivate the MAPKs by dephosphorylation. We are using mouse genetic approaches to study the role of MKP-1 in vivo.
Integration of PTPs and calcium signaling pathways.
Calcium is an important mediator of intracellular signal transduction. We are interested in how PTPs integrate with calcium signaling pathways in the cytoplasm and nucleus. In order to study the integrated role of PTPs and calcium in the nucleus and cytoplasm, we have developed tools to selectively buffer calcium either in the nucleus or cytoplasm independently. We are employing these targeted calcium buffering tools to determine the compartmental affects of calcium on PTP function.
Extensive Research Description
Cell proliferation, cell differentiation and death are controlled by signaling pathways that are mediated by protein tyrosyl phosphorylation. The net cellular level of protein tyrosyl phosphorylation is regulated by the intrinsic and opposing activities of protein tyrosine kinases and protein tyrosine phosphatases (PTPs). Dysregulation of the net level of tyrosyl phosphorylation causes cancer, diabetes, cardiovascular disease and developmental abnormalities.
We focus our studies on how PTPs regulate cell signaling by controlling protein tyrosyl dephosphorylation. We are also exploring how PTPs participate in the pathogenesis of diseases such as obesity and diabetes, cardiovascular disease and developmental abnormalities. These research efforts are being accomplished by using integrated approaches that encompass cell biology, genetics, protein biochemistry, cell imaging and animal models. Our long-term research goals are to uncover the function of PTPs in order to identify whether these enzymes might serve as novel therapeutic targets for the treatment of human diseases.
Improved Regenerative Myogenesis and Muscular Dystrophy in Mice lacking MKP-5
Shi, H., Verma, M., Zhang, L. Dong, C., Flavell, R.A., and Bennett, A. M. (2013), Improved Regenerative Myogenesis and Muscular Dystrophy in Mice lacking MKP-5, J. Clin. Invest., 123: 2064-2077.
Diversity of mitogen-activated protein kinase phosphatase-1 functions, Cellular and Molecular Life Sciences (CMLS)
Lawan, A., Shi, H., Gratzke, F. and Bennett, A.M. (2013) Diversity of mitogen-activated protein kinase phosphatase-1 functions, Cellular and Molecular Life Sciences (CMLS), 70:223-237.
A Novel Role for SHP-2 in Nutrient-Responsive Control of S6 Kinase 1 Signaling
Mercan, F., Lee, H., Kolli, S. and Bennett, A. M. (2013) A Novel Role for SHP-2 in Nutrient-Responsive Control of S6 Kinase 1 Signaling, Mol. Cell Biol., 33: 293-306.
Loss of MAP Kinase Phosphatase-1 protects from hepatic steatosis by repression of CIDEC/Fat-specific protein 27
Roth Flach, R.J., Zhang, and Bennett, A.M. (2011) Loss of MAP Kinase Phosphatase-1 protects from hepatic steatosis by repression of CIDEC/Fat-specific protein 27, J. Biol. Chem., 286: 22195-22202.
MAPK phosphatase-1 facilitates the loss of oxidative myofibers associated with obesity in mice
Roth, R.J., Le, A., Zhang, L., Samuel, V.T., Shulman, J., and Bennett, A.M., (2009) MAPK phosphatase-1 facilitates the loss of oxidative myofibers associated with obesity in mice. J. Clin. Invest., 119:3817-29.
Full List of PubMed Publications
- Tzouvelekis A, Yu G, Lino Cardenas CL, Herazo-Maya JD, Wang R, Woolard T, Zhang Y, Sakamoto K, Lee H, Yi JS, DeIuliis G, Xylourgidis N, Ahangari F, Lee PJ, Aidinis V, Herzog EL, Homer R, Bennett AM, Kaminski N: SH2 Domain-Containing Phosphatase-2 Is a Novel Antifibrotic Regulator in Pulmonary Fibrosis. Am J Respir Crit Care Med. 2017 Feb 15. PMID: 27736153
- Lee H, Min K, Yi JS, Shi H, Chang W, Jackson L, Bennett AM: A phosphoproteomic screen identifies a guanine-nucleotide exchange factor for Rab3A as a MAP kinase phosphatase-5-regulated MAP kinase target in IL-6 secretion and myogenesis. J Biol Chem. 2017 Jan 17; 2017 Jan 17. PMID: 28096466
- Yi JS, Huang Y, Kwaczala AT, Kuo IY, Ehrlich BE, Campbell SG, Giordano FJ, Bennett AM: Low-dose dasatinib rescues cardiac function in Noonan syndrome. JCI Insight. 2016 Dec 8; 2016 Dec 8. PMID: 27942593
- Shi H, Gatzke F, Molle JM, Lee HB, Helm ET, Oldham JJ, Zhang L, Gerrard DE, Bennett AM: Mice lacking MKP-1 and MKP-5 Reveal Hierarchical Regulation of Regenerative Myogenesis. J Stem Cell Regen Biol. 2015 Nov 12. PMID: 27064463
- Lee H, Bennett AM: Identification of receptor protein tyrosine phosphatases (RPTPs) as regulators of receptor tyrosine kinases (RTKs) using an RPTP siRNA-RTK substrate screen. Methods Mol Biol. 2015. PMID: 25319894
- Lawan A, Zhang L, Gatzke F, Min K, Jurczak MJ, Al-Mutairi M, Richter P, Camporez JP, Couvillon A, Pesta D, Roth Flach RJ, Shulman GI, Bennett AM: Hepatic mitogen-activated protein kinase phosphatase 1 selectively regulates glucose metabolism and energy homeostasis. Mol Cell Biol. 2015 Jan; 2014 Oct 13. PMID: 25312648
- Lee H, Yi JS, Lawan A, Min K, Bennett AM: Mining the function of protein tyrosine phosphatases in health and disease. Semin Cell Dev Biol. 2015 Jan; 2014 Sep 26. PMID: 25263013
- Feriod CN, Nguyen L, Jurczak MJ, Kruglov EA, Nathanson MH, Shulman GI, Bennett AM, Ehrlich BE: Inositol 1,4,5-trisphosphate receptor type II (InsP3R-II) is reduced in obese mice, but metabolic homeostasis is preserved in mice lacking InsP3R-II. Am J Physiol Endocrinol Metab. 2014 Dec 1; 2014 Oct 14. PMID: 25315698
- Paardekooper Overman J, Yi JS, Bonetti M, Soulsby M, Preisinger C, Stokes MP, Hui L, Silva JC, Overvoorde J, Giansanti P, Heck AJ, Kontaridis MI, den Hertog J, Bennett AM: PZR coordinates Shp2 Noonan and LEOPARD syndrome signaling in zebrafish and mice. Mol Cell Biol. 2014 Aug; 2014 May 27. PMID: 24865967
- Kallen AN, Zhou XB, Xu J, Qiao C, Ma J, Yan L, Lu L, Liu C, Yi JS, Zhang H, Min W, Bennett AM, Gregory RI, Ding Y, Huang Y: The imprinted H19 lncRNA antagonizes let-7 microRNAs. Mol Cell. 2013 Oct 10; 2013 Sep 19. PMID: 24055342
- Shi H, Verma M, Zhang L, Dong C, Flavell RA, Bennett AM: Improved regenerative myogenesis and muscular dystrophy in mice lacking Mkp5. J Clin Invest. 2013 May; 2013 Apr 1. PMID: 23543058
- Lee H, Bennett AM: Receptor protein tyrosine phosphatase-receptor tyrosine kinase substrate screen identifies EphA2 as a target for LAR in cell migration. Mol Cell Biol. 2013 Apr; 2013 Jan 28. PMID: 23358419
- Lawan A, Shi H, Gatzke F, Bennett AM: Diversity and specificity of the mitogen-activated protein kinase phosphatase-1 functions. Cell Mol Life Sci. 2013 Jan; 2012 Jun 14. PMID: 22695679
- Mercan F, Lee H, Kolli S, Bennett AM: Novel role for SHP-2 in nutrient-responsive control of S6 kinase 1 signaling. Mol Cell Biol. 2013 Jan; 2012 Nov 5. PMID: 23129808
- Ruan HB, Han X, Li MD, Singh JP, Qian K, Azarhoush S, Zhao L, Bennett AM, Samuel VT, Wu J, Yates JR 3rd, Yang X: O-GlcNAc transferase/host cell factor C1 complex regulates gluconeogenesis by modulating PGC-1α stability. Cell Metab. 2012 Aug 8. PMID: 22883232
- Diano S, Liu ZW, Jeong JK, Dietrich MO, Ruan HB, Kim E, Suyama S, Kelly K, Gyengesi E, Arbiser JL, Belsham DD, Sarruf DA, Schwartz MW, Bennett AM, Shanabrough M, Mobbs CV, Yang X, Gao XB, Horvath TL: Peroxisome proliferation-associated control of reactive oxygen species sets melanocortin tone and feeding in diet-induced obesity. Nat Med. 2011 Aug 28; 2011 Aug 28. PMID: 21873987
- Flach RJ, Qin H, Zhang L, Bennett AM: Loss of mitogen-activated protein kinase phosphatase-1 protects from hepatic steatosis by repression of cell death-inducing DNA fragmentation factor A (DFFA)-like effector C (CIDEC)/fat-specific protein 27. J Biol Chem. 2011 Jun 24; 2011 Apr 26. PMID: 21521693
- Roth Flach RJ, Bennett AM: Mitogen-activated protein kinase phosphatase-1 - a potential therapeutic target in metabolic disease. Expert Opin Ther Targets. 2010 Dec. PMID: 21058921
- Shi H, Boadu E, Mercan F, Le AM, Flach RJ, Zhang L, Tyner KJ, Olwin BB, Bennett AM: MAP kinase phosphatase-1 deficiency impairs skeletal muscle regeneration and exacerbates muscular dystrophy. FASEB J. 2010 Aug; 2010 Apr 6. PMID: 20371627
- Mercan F, Bennett AM: Analysis of protein tyrosine phosphatases and substrates. Curr Protoc Mol Biol. 2010 Jul. PMID: 20583096
- Flach RJ, Bennett AM: MAP kinase phosphatase-1--a new player at the nexus between sarcopenia and metabolic disease. Aging (Albany NY). 2010 Apr 6; 2010 Apr 6. PMID: 20375469
- Roth RJ, Le AM, Zhang L, Kahn M, Samuel VT, Shulman GI, Bennett AM: MAPK phosphatase-1 facilitates the loss of oxidative myofibers associated with obesity in mice. J Clin Invest. 2009 Dec; 2009 Nov 16. PMID: 19920356
- Carlson J, Cui W, Zhang Q, Xu X, Mercan F, Bennett AM, Vignery A: Role of MKP-1 in osteoclasts and bone homeostasis. Am J Pathol. 2009 Oct; 2009 Sep 17. PMID: 19762714
- Soulsby M, Bennett AM: Physiological signaling specificity by protein tyrosine phosphatases. Physiology (Bethesda). 2009 Oct. PMID: 19815854
- Yu L, Min W, He Y, Qin L, Zhang H, Bennett AM, Chen H: JAK2 and SHP2 reciprocally regulate tyrosine phosphorylation and stability of proapoptotic protein ASK1. J Biol Chem. 2009 May 15; 2009 Mar 13. PMID: 19287004
- Eminaga S, Bennett AM: Noonan syndrome-associated SHP-2/Ptpn11 mutants enhance SIRPalpha and PZR tyrosyl phosphorylation and promote adhesion-mediated ERK activation. J Biol Chem. 2008 May 30; 2008 Mar 31. PMID: 18378677
- Gomes DA, Rodrigues MA, Leite MF, Gomez MV, Varnai P, Balla T, Bennett AM, Nathanson MH: c-Met must translocate to the nucleus to initiate calcium signals. J Biol Chem. 2008 Feb 15; 2007 Dec 11. PMID: 18073207
- Rodrigues MA, Gomes DA, Leite MF, Grant W, Zhang L, Lam W, Cheng YC, Bennett AM, Nathanson MH: Nucleoplasmic calcium is required for cell proliferation. J Biol Chem. 2007 Jun 8; 2007 Apr 9. PMID: 17420246
- Zito CI, Qin H, Blenis J, Bennett AM: SHP-2 regulates cell growth by controlling the mTOR/S6 kinase 1 pathway. J Biol Chem. 2007 Mar 9; 2007 Jan 17. PMID: 17229738
- Fornaro M, Burch PM, Yang W, Zhang L, Hamilton CE, Kim JH, Neel BG, Bennett AM: SHP-2 activates signaling of the nuclear factor of activated T cells to promote skeletal muscle growth. J Cell Biol. 2006 Oct 9; 2006 Oct 2. PMID: 17015617
- Wu JJ, Roth RJ, Anderson EJ, Hong EG, Lee MK, Choi CS, Neufer PD, Shulman GI, Kim JK, Bennett AM: Mice lacking MAP kinase phosphatase-1 have enhanced MAP kinase activity and resistance to diet-induced obesity. Cell Metab. 2006 Jul. PMID: 16814733
- Chi H, Barry SP, Roth RJ, Wu JJ, Jones EA, Bennett AM, Flavell RA: Dynamic regulation of pro- and anti-inflammatory cytokines by MAPK phosphatase 1 (MKP-1) in innate immune responses. Proc Natl Acad Sci U S A. 2006 Feb 14; 2006 Feb 6. PMID: 16461893
- Uhlén P, Burch PM, Zito CI, Estrada M, Ehrlich BE, Bennett AM: Gain-of-function/Noonan syndrome SHP-2/Ptpn11 mutants enhance calcium oscillations and impair NFAT signaling. Proc Natl Acad Sci U S A. 2006 Feb 14; 2006 Feb 3. PMID: 16461457
- Park SY, Cho YR, Finck BN, Kim HJ, Higashimori T, Hong EG, Lee MK, Danton C, Deshmukh S, Cline GW, Wu JJ, Bennett AM, Rothermel B, Kalinowski A, Russell KS, Kim YB, Kelly DP, Kim JK: Cardiac-specific overexpression of peroxisome proliferator-activated receptor-alpha causes insulin resistance in heart and liver. Diabetes. 2005 Sep. PMID: 16123338
- Wu JJ, Zhang L, Bennett AM: The noncatalytic amino terminus of mitogen-activated protein kinase phosphatase 1 directs nuclear targeting and serum response element transcriptional regulation. Mol Cell Biol. 2005 Jun. PMID: 15899879
- Wu JJ, Bennett AM: Essential role for mitogen-activated protein (MAP) kinase phosphatase-1 in stress-responsive MAP kinase and cell survival signaling. J Biol Chem. 2005 Apr 22; 2005 Feb 17. PMID: 15722358
- Chauvet V, Tian X, Husson H, Grimm DH, Wang T, Hiesberger T, Igarashi P, Bennett AM, Ibraghimov-Beskrovnaya O, Somlo S, Caplan MJ: Mechanical stimuli induce cleavage and nuclear translocation of the polycystin-1 C terminus. J Clin Invest. 2004 Nov. PMID: 15545994
- Kolli S, Zito CI, Mossink MH, Wiemer EA, Bennett AM: The major vault protein is a novel substrate for the tyrosine phosphatase SHP-2 and scaffold protein in epidermal growth factor signaling. J Biol Chem. 2004 Jul 9; 2004 May 7. PMID: 15133037
- Kontaridis MI, Eminaga S, Fornaro M, Zito CI, Sordella R, Settleman J, Bennett AM: SHP-2 positively regulates myogenesis by coupling to the Rho GTPase signaling pathway. Mol Cell Biol. 2004 Jun. PMID: 15169898
- Ivins Zito C, Kontaridis MI, Fornaro M, Feng GS, Bennett AM: SHP-2 regulates the phosphatidylinositide 3'-kinase/Akt pathway and suppresses caspase 3-mediated apoptosis. J Cell Physiol. 2004 May. PMID: 15040005
- Pusl T, Wu JJ, Zimmerman TL, Zhang L, Ehrlich BE, Berchtold MW, Hoek JB, Karpen SJ, Nathanson MH, Bennett AM: Epidermal growth factor-mediated activation of the ETS domain transcription factor Elk-1 requires nuclear calcium. J Biol Chem. 2002 Jul 26; 2002 Apr 23. PMID: 11971908
- Kontaridis MI, Liu X, Zhang L, Bennett AM: Role of SHP-2 in fibroblast growth factor receptor-mediated suppression of myogenesis in C2C12 myoblasts. Mol Cell Biol. 2002 Jun. PMID: 11997521