XB-ART-44719
Bioorg Med Chem
2012 Mar 15;206:1869-77. doi: 10.1016/j.bmc.2011.12.044.
Show Gene links
Show Anatomy links
Screening for small molecule inhibitors of embryonic pathways: sometimes you gotta crack a few eggs.
Hang BI
,
Thorne CA
,
Robbins DJ
,
Huppert SS
,
Lee LA
,
Lee E
.
???displayArticle.abstract???
Extract prepared from Xenopus eggs represents a cell-free system that has been shown to recapitulate a multitude of cellular processes, including cell cycle regulation, DNA replication/repair, and cytoskeletal dynamics. In addition, this system has been used to successfully reconstitute the Wnt pathway. Xenopus egg extract, which can be biochemically manipulated, offers an ideal medium in which small molecule screening can be performed in near native milieu. Thus, the use of Xenopus egg extract for small molecule screening represents an ideal bridge between targeted and phenotypic screening approaches. This review focuses on the use of this system for small molecules modulators of major signal transduction pathways (Notch, Hedgehog, and Wnt) that are critical for the development of the early Xenopus embryo. We describe the properties of Xenopus egg extract and our own high throughput screen for small molecules that modulate the Wnt pathway using this cell-free system. We propose that Xenopus egg extract could similarly be adapted for screening for modulators of the Notch and Hedgehog pathways.
???displayArticle.pubmedLink??? 22261025
???displayArticle.pmcLink??? PMC3298638
???displayArticle.link??? Bioorg Med Chem
???displayArticle.grants??? [+]
Species referenced: Xenopus laevis
Genes referenced: notch1 shh
References [+] :
Andersson,
Notch signaling: simplicity in design, versatility in function.
2011, Pubmed
Andersson, Notch signaling: simplicity in design, versatility in function. 2011, Pubmed
Arias, Initiation of DNA replication in Xenopus egg extracts. 2004, Pubmed , Xenbase
Barker, Mining the Wnt pathway for cancer therapeutics. 2006, Pubmed
Barton, The erythroid protein cGATA-1 functions with a stage-specific factor to activate transcription of chromatin-assembled beta-globin genes. 1993, Pubmed
Beachy, Tissue repair and stem cell renewal in carcinogenesis. 2004, Pubmed
Blow, Replication of purified DNA in Xenopus egg extract is dependent on nuclear assembly. 1990, Pubmed , Xenbase
Blow, Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs. 1986, Pubmed , Xenbase
Cameron, Secrets of actin-based motility revealed by a bacterial pathogen. 2000, Pubmed
Chan, In vitro study of nuclear assembly and nuclear import using Xenopus egg extracts. 2006, Pubmed , Xenbase
Clevers, Wnt/beta-catenin signaling in development and disease. 2006, Pubmed
Cselenyi, LRP6 transduces a canonical Wnt signal independently of Axin degradation by inhibiting GSK3's phosphorylation of beta-catenin. 2008, Pubmed , Xenbase
Dabauvalle, Assembly of nuclear pore complexes in Xenopus egg extract. 1991, Pubmed , Xenbase
Dahmane, Activation of the transcription factor Gli1 and the Sonic hedgehog signalling pathway in skin tumours. 1997, Pubmed , Xenbase
Daniels, Beta-catenin directly displaces Groucho/TLE repressors from Tcf/Lef in Wnt-mediated transcription activation. 2005, Pubmed
DeMoe, A high throughput, whole cell screen for small molecule inhibitors of the mitotic spindle checkpoint identifies OM137, a novel Aurora kinase inhibitor. 2009, Pubmed
Deming, Study of apoptosis in vitro using the Xenopus egg extract reconstitution system. 2006, Pubmed , Xenbase
Desai, Microtubule polymerization dynamics. 1997, Pubmed
Desai, The use of Xenopus egg extracts to study mitotic spindle assembly and function in vitro. 1999, Pubmed , Xenbase
Dick, Looking ahead in cancer stem cell research. 2009, Pubmed
Finlay, Inhibition of in vitro nuclear transport by a lectin that binds to nuclear pores. 1987, Pubmed , Xenbase
Frank, The therapeutic promise of the cancer stem cell concept. 2010, Pubmed
Garner, Studying the DNA damage response using in vitro model systems. 2009, Pubmed , Xenbase
Glotzer, Cyclin is degraded by the ubiquitin pathway. 1991, Pubmed , Xenbase
Green, Basal cell carcinoma development is associated with induction of the expression of the transcription factor Gli-1. 1998, Pubmed
Hahn, Mutations of the human homolog of Drosophila patched in the nevoid basal cell carcinoma syndrome. 1996, Pubmed
Hannak, Investigating mitotic spindle assembly and function in vitro using Xenopus laevis egg extracts. 2006, Pubmed , Xenbase
Heald, Self-organization of microtubules into bipolar spindles around artificial chromosomes in Xenopus egg extracts. 1996, Pubmed , Xenbase
Ho, Toca-1 mediates Cdc42-dependent actin nucleation by activating the N-WASP-WIP complex. 2004, Pubmed , Xenbase
Hoppler, Studying Wnt signaling in Xenopus. 2008, Pubmed , Xenbase
Hubbard, sel-10, a negative regulator of lin-12 activity in Caenorhabditis elegans, encodes a member of the CDC4 family of proteins. 1997, Pubmed
Ingham, Mechanisms and functions of Hedgehog signalling across the metazoa. 2011, Pubmed
Johnson, Human homolog of patched, a candidate gene for the basal cell nevus syndrome. 1996, Pubmed
Katoh, Hedgehog target genes: mechanisms of carcinogenesis induced by aberrant hedgehog signaling activation. 2009, Pubmed
King, A 20S complex containing CDC27 and CDC16 catalyzes the mitosis-specific conjugation of ubiquitin to cyclin B. 1995, Pubmed , Xenbase
Kinzler, Lessons from hereditary colorectal cancer. 1996, Pubmed
Kinzler, Identification of FAP locus genes from chromosome 5q21. 1991, Pubmed
Kinzler, Identification of an amplified, highly expressed gene in a human glioma. 1987, Pubmed
Kofron, Wnt11/beta-catenin signaling in both oocytes and early embryos acts through LRP6-mediated regulation of axin. 2007, Pubmed , Xenbase
Kopan, The canonical Notch signaling pathway: unfolding the activation mechanism. 2009, Pubmed
Kornbluth, Analysis of the cell cycle using Xenopus egg extracts. 2006, Pubmed , Xenbase
Kwok, A small-molecule screen in C. elegans yields a new calcium channel antagonist. 2006, Pubmed
Lee, The roles of APC and Axin derived from experimental and theoretical analysis of the Wnt pathway. 2003, Pubmed , Xenbase
Lindsay, Target discovery. 2003, Pubmed
Logan, The Wnt signaling pathway in development and disease. 2004, Pubmed
Lohka, Formation in vitro of sperm pronuclei and mitotic chromosomes induced by amphibian ooplasmic components. 1983, Pubmed , Xenbase
Lohka, Roles of cytosol and cytoplasmic particles in nuclear envelope assembly and sperm pronuclear formation in cell-free preparations from amphibian eggs. 1984, Pubmed , Xenbase
Lohka, Purification of maturation-promoting factor, an intracellular regulator of early mitotic events. 1988, Pubmed , Xenbase
Ma, Corequirement of specific phosphoinositides and small GTP-binding protein Cdc42 in inducing actin assembly in Xenopus egg extracts. 1998, Pubmed , Xenbase
MacDonald, Wnt/beta-catenin signaling: components, mechanisms, and diseases. 2009, Pubmed , Xenbase
Mao, Low-density lipoprotein receptor-related protein-5 binds to Axin and regulates the canonical Wnt signaling pathway. 2001, Pubmed
Maresca, Methods for studying spindle assembly and chromosome condensation in Xenopus egg extracts. 2006, Pubmed , Xenbase
Matthews, A highly efficient, cell-free translation/translocation system prepared from Xenopus eggs. 1991, Pubmed , Xenbase
Meier, The role of lamin LIII in nuclear assembly and DNA replication, in cell-free extracts of Xenopus eggs. 1991, Pubmed , Xenbase
Minshull, Translation of cyclin mRNA is necessary for extracts of activated xenopus eggs to enter mitosis. 1989, Pubmed , Xenbase
Mitchison, Towards a pharmacological genetics. 1994, Pubmed
Mitchison, Dynamic instability of microtubule growth. , Pubmed
Murray, Cyclin synthesis drives the early embryonic cell cycle. 1989, Pubmed , Xenbase
Murray, The role of cyclin synthesis and degradation in the control of maturation promoting factor activity. 1989, Pubmed , Xenbase
Murray, Cell cycle extracts. 1991, Pubmed
Newmeyer, Cell-free apoptosis in Xenopus egg extracts: inhibition by Bcl-2 and requirement for an organelle fraction enriched in mitochondria. 1994, Pubmed , Xenbase
Newmeyer, In vitro transport of a fluorescent nuclear protein and exclusion of non-nuclear proteins. 1986, Pubmed , Xenbase
Newport, A lamin-independent pathway for nuclear envelope assembly. 1990, Pubmed , Xenbase
Newport, A major developmental transition in early Xenopus embryos: I. characterization and timing of cellular changes at the midblastula stage. 1982, Pubmed , Xenbase
Newport, A major developmental transition in early Xenopus embryos: II. Control of the onset of transcription. 1982, Pubmed , Xenbase
Newport, Nuclear reconstitution in vitro: stages of assembly around protein-free DNA. 1987, Pubmed , Xenbase
Newport, Disassembly of the nucleus in mitotic extracts: membrane vesicularization, lamin disassembly, and chromosome condensation are independent processes. 1987, Pubmed , Xenbase
Nishisho, Mutations of chromosome 5q21 genes in FAP and colorectal cancer patients. 1991, Pubmed
Nutt, Metabolic regulation of oocyte cell death through the CaMKII-mediated phosphorylation of caspase-2. 2005, Pubmed , Xenbase
O'Neil, FBW7 mutations in leukemic cells mediate NOTCH pathway activation and resistance to gamma-secretase inhibitors. 2007, Pubmed
Oberg, The Notch intracellular domain is ubiquitinated and negatively regulated by the mammalian Sel-10 homolog. 2001, Pubmed
Ogden, Regulation of Hedgehog signaling: a complex story. 2004, Pubmed
Overington, How many drug targets are there? 2006, Pubmed
Peterson, Discovery and use of small molecules for probing biological processes in zebrafish. 2004, Pubmed
Peterson, A chemical inhibitor of N-WASP reveals a new mechanism for targeting protein interactions. 2001, Pubmed , Xenbase
Pinto, Wnt control of stem cells and differentiation in the intestinal epithelium. 2005, Pubmed
Pomerening, Systems-level dissection of the cell-cycle oscillator: bypassing positive feedback produces damped oscillations. 2005, Pubmed , Xenbase
Ranganathan, Notch signalling in solid tumours: a little bit of everything but not all the time. 2011, Pubmed
Robbins, Hedgehogs: la dolce vita. Workshop on Hedgehog-Gli Signaling in Cancer and Stem Cells. 2007, Pubmed
Rohatgi, Mechanism of N-WASP activation by CDC42 and phosphatidylinositol 4, 5-bisphosphate. 2000, Pubmed , Xenbase
Rosenblatt, Xenopus actin depolymerizing factor/cofilin (XAC) is responsible for the turnover of actin filaments in Listeria monocytogenes tails. 1997, Pubmed , Xenbase
Rubin, Targeting the Hedgehog pathway in cancer. 2006, Pubmed
Salic, Control of beta-catenin stability: reconstitution of the cytoplasmic steps of the wnt pathway in Xenopus egg extracts. 2000, Pubmed , Xenbase
Saraswati, Pyrvinium, a potent small molecule Wnt inhibitor, promotes wound repair and post-MI cardiac remodeling. 2010, Pubmed
Sawin, Mitotic spindle assembly by two different pathways in vitro. 1991, Pubmed , Xenbase
Sawin, Poleward microtubule flux mitotic spindles assembled in vitro. 1991, Pubmed , Xenbase
Shennan, Xenopus egg extracts: a model system to study proprotein convertases. 2006, Pubmed , Xenbase
Shirasu, Microtubule dynamics in Xenopus egg extracts. 1999, Pubmed , Xenbase
Spencer, Measuring and modeling apoptosis in single cells. 2011, Pubmed
Su, Multiple intestinal neoplasia caused by a mutation in the murine homolog of the APC gene. 1992, Pubmed
Swinney, How were new medicines discovered? 2011, Pubmed
Theriot, Involvement of profilin in the actin-based motility of L. monocytogenes in cells and in cell-free extracts. 1994, Pubmed , Xenbase
Thorne, Small-molecule inhibition of Wnt signaling through activation of casein kinase 1α. 2010, Pubmed , Xenbase
Thorne, A biochemical screen for identification of small-molecule regulators of the Wnt pathway using Xenopus egg extracts. 2011, Pubmed , Xenbase
Tolwinski, Wg/Wnt signal can be transmitted through arrow/LRP5,6 and Axin independently of Zw3/Gsk3beta activity. 2003, Pubmed
Toyoda, Characterization of RNA polymerase II-dependent transcription in Xenopus extracts. 1992, Pubmed , Xenbase
Tutter, Chromosomal DNA replication in a soluble cell-free system derived from Xenopus eggs. 2006, Pubmed , Xenbase
Verma, Ubistatins inhibit proteasome-dependent degradation by binding the ubiquitin chain. 2004, Pubmed , Xenbase
Walter, Regulated chromosomal DNA replication in the absence of a nucleus. 1998, Pubmed , Xenbase
Walter, Initiation of eukaryotic DNA replication: origin unwinding and sequential chromatin association of Cdc45, RPA, and DNA polymerase alpha. 2000, Pubmed , Xenbase
Weng, Activating mutations of NOTCH1 in human T cell acute lymphoblastic leukemia. 2004, Pubmed
Westhoff, Alterations of the Notch pathway in lung cancer. 2009, Pubmed
Wignall, Identification of a novel protein regulating microtubule stability through a chemical approach. 2004, Pubmed , Xenbase
Wolfe, gamma-Secretase in biology and medicine. 2009, Pubmed
Wu, SEL-10 is an inhibitor of notch signaling that targets notch for ubiquitin-mediated protein degradation. 2001, Pubmed
Yauch, Smoothened mutation confers resistance to a Hedgehog pathway inhibitor in medulloblastoma. 2009, Pubmed
Yeh, Chemical genetics: adding to the developmental biology toolbox. 2003, Pubmed