A Zebrafish Chemical Suppressor Screening Identifies Small Molecule Inhibitors of the Wnt/β-catenin Pathway

SummaryGenetic screening for suppressor mutants has been successfully used to identify important signaling regulators. Using an analogy to genetic suppressor screening, we developed a chemical suppressor screening method to identify inhibitors of the Wnt/β-catenin signaling pathway. We used zebrafish embryos in which chemically induced β-catenin accumulation led to an “eyeless” phenotype and conducted a pilot screening for compounds that restored eye development. This approach allowed us to identify geranylgeranyltransferase inhibitor 286 (GGTI-286), a geranylgeranyltransferase (GGTase) inhibitor. Our follow-up studies showed that GGTI-286 reduces nuclear localization of β-catenin and transcription dependent on β-catenin/T cell factor in mammalian cells. In addition to pharmacological inhibition, GGTase gene knockdown also attenuates the nuclear function of β-catenin. Overall, we validate our chemical suppressor screening as a method for identifying Wnt/β-catenin pathway inhibitors and implicate GGTase as a potential therapeutic target for Wnt-activated cancers

The Dichotomous Nucleon: Some Radical Conjectures for the Large Nc Limit

We discuss some problems with the large Nc approximation for nucleons which arise if the axial coupling of the nucleon to pions is large, g_A \sim Nc. While g_A \sim Nc in non-relativistic quark and Skyrme models, it has been suggested that Skyrmions may collapse to a small size, r \sim 1/f_pi \sim Lambda_QCD^{-1}/sqrt(Nc). (This is also the typical scale over which the string vertex moves in a string vertex model of the baryon.) We concentrate on the case of two flavors, where we suggest that to construct a nucleon with a small axial coupling, that most quarks are bound into colored diquark pairs, which have zero spin and isospin. For odd Nc, this leaves one unpaired quark, which carries the spin and isospin of the nucleon. If the unpaired quark is in a spatial wavefunction orthogonal to the wavefunctions of the scalar diquarks, then up to logarithms of Nc, the unpaired quark only costs an energy \sim Lambda_QCD. This naturally gives g_A \sim 1 and has other attractive features. In nature, the wavefunctions of the paired and unpaired quarks might only be approximately orthogonal; then g_A depends weakly upon Nc. This dichotomy in wave functions could arise if the unpaired quark orbits at a size which is parametrically large in comparison to that of the diquarks. We discuss possible tests of these ideas from numerical simulations on the lattice, for two flavors and three and five colors; the extension of our ideas to more than three or more flavors is not obvious, though.Comment: Published version in Nucl. Phys.

Novel Approach to Designing Primers for Identification and Distinction of the Human Pathogenic Fungi Coccidioides immitis and Coccidioides posadasii by PCR Amplification

We developed a pair of primers that specifically identifies Coccidioides species, etiologic agents of the human fungal disease coccidioidomycosis. These primers could be used for distinguishing Coccidioides immitis and Coccidioides posadasii by simply comparing the amplicon sizes on an agarose gel

Tcc1p, a Novel Protein Containing the Tetratricopeptide Repeat Motif, Interacts with Tup1p To Regulate Morphological Transition and Virulence in Candida albicans

The transcriptional factor CaTup1p represses many genes involved in intracellular processes, including the yeast-hypha transition, in the human fungal pathogen Candida albicans. Using tandem affinity purification technology, we identified a novel protein that interacts with CaTup1p, named Tcc1p (Tup1p complex component). Tcc1p is a C. albicans-specific protein with a 736-amino-acid polypeptide with four tetratricopeptide repeat (TPR) motifs in the N-terminal portion. Tcc1p formed a protein complex with CaTup1p via the TPR domain of Tcc1p, independently of CaSsn6p-CaTup1p The tcc1Δ disruptant showed filamentous growth under conditions inducing the yeast form, as is true of the Catup1Δ mutant. Consistent with this result, the common set of hypha-specific genes was negatively regulated by both TCC1 and CaTUP1. These observations will provide new insights into CaTup1p-dependent transcriptional gene regulation in C. albicans

Small molecules inhibiting the nuclear localization of YAP/TAZ for chemotherapeutics and chemosensitizers against breast cancers

YAP and TAZ oncoproteins confer malignancy and drug resistance to various cancer types. We screened for small molecules that inhibit the nuclear localization of YAP/TAZ. Dasatinib, statins and pazopanib inhibited the nuclear localization and target gene expression of YAP and TAZ. All three drugs induced phosphorylation of YAP and TAZ, and pazopanib induced proteasomal degradation of YAP/TAZ. The sensitivities to these drugs are correlated with dependence on YAP/TAZ in breast cancer cell lines. Combinations of these compounds with each other or with other anti‐cancer drugs efficiently reduced cell proliferation of YAP/TAZ‐dependent breast cancer cells. These results suggest that these drugs can be therapeutics and chemosensitizers for YAP/TAZ‐dependent breast cancers