A small molecule accelerates neuronal differentiation in the adult rat

Wurdak, Heiko; Zhu, Shoutian; Min, Kyung Hoon; Aimone, Lindsey J.; Lairson, Luke L.; Watson, James P.; Chopiuk, Gregory; Demas, Jay; Charette, Bradley D.; Halder, Rajkumar; Weerapana, Eranthie; Cravatt, Benjamin F.; Cline, Hollis T.; Peters, Eric C.; Zhang, Jay; Walker, John R.; Wu, Chunlei; Chang, Jonathan; Tuntland, Tove; Cho, Charles Y.; Schultz, Peter G.

doi:10.1073/pnas.1010300107

Cited by 106 publications

(114 citation statements)

References 44 publications

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“…A similar role was also recently reported for TACC3 and ARNT2 in neuronal progenitor cells. Disruption of the TACC3/ARNT2 interaction by a small molecule inhibitor accelerated differentiation within neural progenitor cells (39), suggesting that the ARNT PAS-B mediated interaction described here may play an important role in mediating cell fate decisions by several types of stem cell populations.…”

Section: Mutations In Arnt Pas-b Perturb Coactivator Binding and Hifmentioning

confidence: 81%

Coactivators necessary for transcriptional output of the hypoxia inducible factor, HIF, are directly recruited by ARNT PAS-B

Partch

Gardner

2011

Proc. Natl. Acad. Sci. U.S.A.

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Hypoxia-inducible factor (HIF) is the key transcriptional effector of the hypoxia response in eukaryotes, coordinating the expression of genes involved in oxygen transport, glycolysis, and angiogenesis to promote adaptation to low oxygen levels. HIF is a basic helixloop-helix (bHLH)-PAS (PER-ARNT-SIM) heterodimer composed of an oxygen-labile HIF-α subunit and a constitutively expressed aryl hydrocarbon receptor nuclear translocator (ARNT) subunit, which dimerize via basic helix-loop-helix and PAS domains, and recruit coactivators via HIF-α C-terminal transactivation domains. Here we demonstrate that the ARNT PAS-B domain provides an additional recruitment site by binding the coactivator transforming acidic coiled-coil 3 (TACC3) in a step necessary for transcriptional responses to hypoxia. Structural insights from NMR spectroscopy illustrate how this PAS domain simultaneously mediates interactions with HIF-α and TACC3. Finally, mutations on ARNT PAS-B modulate coactivator selectivity and target gene induction by HIF in vivo, demonstrating a bifunctional role for transcriptional regulation by PAS domains within bHLH-PAS transcription factors. transcriptional coactivators | protein/protein interactions | bifunctional interactionsA ryl hydrocarbon receptor nuclear translocator (ARNT) is the obligate heterodimeric partner for the basic helix-loop-helix (bHLH)-PAS (PER-ARNT-SIM) proteins aryl hydrocarbon receptor (AhR) and hypoxia-inducible factor-α (HIF-α), which serve as environmental sensors for xenobiotics and hypoxia, respectively (1). bHLH-PAS heterodimers are dependent on intersubunit contacts between the basic bHLH and tandem PAS domains (2-4). The second of two PAS domains, PAS-B, plays a critical role in maintaining the stability of this complex, given that mutations in HIF-2α PAS-B disrupt HIF-α/ARNT interactions and decrease transactivation in vivo (3, 4). Therefore, our current model of bHLH-PAS heterodimer architecture is based on nucleation of the core transcription factor complex by bHLH and PAS domains, leaving C-terminal transactivation domains (TADs) to recruit coactivator proteins that are required for gene regulation (Fig. 1A).Further study of HIF TADs reveals that not all are essential for HIF function. In particular, deletion of the putative ARNT C-terminal TAD has a minimal effect on transactivation of endogenous targets (5-7), whereas deletion of the two HIF-α TADs (N-TAD and C-TAD) eliminates hypoxia-induced transactivation (8). Consequently, study of HIF transcriptional regulation has focused on the HIF-α TADs, identifying the C-TAD as the primary site of recruitment for p300/CBP (9) and the N-TAD as a determining factor in the distinctive profiles of target gene induction by . Selectivity is mediated in part by the recruitment of different coactivators by the N-TADs of the two HIF-α isoforms, building on an emerging theme that transcriptional coregulators and promoter context influence the specificity of gene induction by transcription factors (11,12). Domain-swapping studies have shown tha...

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Section: Mutations In Arnt Pas-b Perturb Coactivator Binding and Hifmentioning

confidence: 81%

Coactivators necessary for transcriptional output of the hypoxia inducible factor, HIF, are directly recruited by ARNT PAS-B

Partch

Gardner

2011

Proc. Natl. Acad. Sci. U.S.A.

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“…The identity of all cell lines was validated by short tandem repeat (STR) analysis at multiple points during the study. KHS101 was a kind gift from Peter Schultz (44). Additional KHS101 was synthesized by the UNC Center for Integrative Chemical Biology and Drug Discovery.…”

Section: Methodsmentioning

confidence: 99%

“…The restricted expression pattern of this cohort of gametogenic proteins and their requirement for deflecting mitotic perturbation suggest that they could represent unique entry points to enhance paclitaxel sensitivity in tumor cells. The recent discovery of a TACC3 inhibitor, KHS101, affords the opportunity to evaluate the therapeutic potential of targeting a reactivated gametogenic protein to enhance sensitivity to the first-line chemotherapeutic paclitaxel (44). To evaluate this possibility, we exposed H1155 cells to a combination of KHS101 and paclitaxel at doses with little or no single-agent activity.…”

Section: Depletion Of Testis Proteins Disrupts Bipolar Spindle Formatmentioning

confidence: 99%

Multiple Cancer Testis Antigens Function To Support Tumor Cell Mitotic Fidelity

Cappell

Sinnott

Taus

et al. 2012

Molecular and Cellular Biology

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While the expression of genes that are normally involved in spermatogenesis is frequently detected in tumors, the extent to which these gene products are required for neoplastic behaviors is unclear. To begin to address their functional relevance to tumorigenesis, we identified a cohort of proteins which display synthetic lethality with paclitaxel in non-small-cell lung cancer and whose expression is biased toward testes and tumors. Remarkably, these testis proteins, FMR1NB, NXF2, MAGEA5, FSIP1, and STARD6, are required for accurate chromosome segregation in tumor cells. Their individual depletion enhances the generation of multipolar spindles, increases mitotic transit time, and induces micronucleation in response to an otherwise innocuous dose of paclitaxel. The underlying basis for abnormal mitosis is an alteration in microtubule function, as their depletion increases microtubule cytaster formation and disrupts microtubule stability. Given these observations, we hypothesize that reactivated testis proteins may represent unique tumor cell vulnerabilities which, if targeted, could enhance responsiveness to antimitotic therapy. Indeed, we demonstrate that combining paclitaxel with a small-molecule inhibitor of the gametogenic and tumor cell mitotic protein TACC3 leads to enhanced centrosomal abnormalities, activation of death programs, and loss of anchorage-independent growth. T umor-specific epigenetic alterations allow the activation of a panoply of otherwise silenced genetic programs that can be exploited to promote neoplastic phenotypes. In particular, genes normally involved in development and gametogenesis are frequently found to be reactivated in tumors, affording an opportunity for transformed cells to co-opt primordial cell features to confer oncogenic behaviors such as self-renewal, uncontrolled proliferation, and epithelial-to-mesenchymal transition (EMT) (1, 39). Cancer testis (CT) antigens represent one group of gametogenic proteins whose expression is otherwise biased toward the testes and reactivated in tumor cells (39). Because the testes are an immune privileged site, humoral and/or cellular responses are often elicited to CT antigens in cancer patients. Thus, much focus has surrounded exploiting these antigens as anticancer vaccine targets, while functional roles, if any, for these proteins in supporting tumor cell fitness have not been extensively evaluated. There are over 200 annotated CT antigens, and most have no known role in spermatogenesis or tumorigenesis (39). However, three recent studies implicate members of the CT antigen family in tumorigenic processes, including p53 turnover, centrosome coalescence, mitotic progression, and regulation of retinoic acid signaling (10,12,43). Furthermore, high expression of specific CT antigens in tumors has been correlated with significantly poorer outcomes, suggesting that their reactivation may enhance aggressiveness or chemoresistance (8,12,43). Therefore, uncovering roles for CT antigens and other ectopically expressed gametogenic genes could si...

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“…For infection experiments, cells were seeded at 10 5 cells per well in six-well dishes and infected with the virus 24 h after plating. The isolation and culture of rat hippocampal neural progenitor cells has been described (18,19). The cells were maintained and expanded in an undifferentiated state by culturing on polyornithine (10 μg/mL in water; Sigma) and laminin (5 μg/mL in PBS; Invitrogen) coated plates.…”

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confidence: 99%

Efficient viral delivery system for unnatural amino acid mutagenesis in mammalian cells

Chatterjee

Xiao

Bollong

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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104

132

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Here we report the development of a baculovirus-based delivery system that enables the efficient incorporation of unnatural amino acids into proteins in mammalian cells. We have exploited the large cargo-capacity (>30 kb) and stability of the double-stranded DNA genome of baculovirus to deliver to a variety of cell types all of the components required to genetically incorporate novel amino acids. These include the engineered tRNA/aminoacyl-tRNA synthetase pair and the nonsense mutant of the target gene. Mammalian cell transduction efficiency of baculovirus was significantly improved by incorporating genetic elements from mammalian viruses. Two polyspecific tRNA/aminoacyl-tRNA synthetase pairs were inserted into this expression system, enabling the site-specific incorporation of a variety of unnatural amino acids with novel chemical and biological properties into proteins.genetic code | viral vector | nonstandard amino acid | orthogonal

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A small molecule accelerates neuronal differentiation in the adult rat

Cited by 106 publications

References 44 publications

Coactivators necessary for transcriptional output of the hypoxia inducible factor, HIF, are directly recruited by ARNT PAS-B

Coactivators necessary for transcriptional output of the hypoxia inducible factor, HIF, are directly recruited by ARNT PAS-B

Multiple Cancer Testis Antigens Function To Support Tumor Cell Mitotic Fidelity

Efficient viral delivery system for unnatural amino acid mutagenesis in mammalian cells

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