Structural Basis of ARNT PAS-B Dimerization: Use of a Common Beta-sheet Interface for Hetero- and Homodimerization

Card, Paul B.; Erbel, P.; Gardner, Kevin H.

doi:10.1016/j.jmb.2005.08.043

Cited by 125 publications

(169 citation statements)

References 48 publications

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“…NMR spectrometry revealed that hydrophobic residues located on the solvent-exposed face of a ␤-sheet in the HIF-2␣ PAS-B subdomain mediate heterodimerization with HIF-1␤ (12). The HIF-2␣ PAS-B crystal structure was shown to contain an internal cavity, which accommodated a small molecule that partially disrupted the heterodimerization of isolated HIF-2␣ and HIF-1␤ PAS subdomains in vitro but was not reported to have any effect on heterodimerization of the full length proteins or on HIF-1 transcriptional activity (13).…”

Section: Discussionmentioning

confidence: 99%

RETRACTED: Acriflavine inhibits HIF-1 dimerization, tumor growth, and vascularization

Lee¹,

Zhang²,

Qian³

et al. 2009

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

434

399

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HIF-1 is a heterodimeric transcription factor that mediates adaptive responses to hypoxia and plays critical roles in cancer progression. Using a cell-based screening assay we have identified acriflavine as a drug that binds directly to HIF-1␣ and HIF-2␣ and inhibits HIF-1 dimerization and transcriptional activity. Pretreatment of mice bearing prostate cancer xenografts with acriflavine prevented tumor growth and treatment of mice bearing established tumors resulted in growth arrest. Acriflavine treatment inhibited intratumoral expression of angiogenic cytokines, mobilization of angiogenic cells into peripheral blood, and tumor vascularization. These results provide proof of principle that small molecules can inhibit dimerization of HIF-1 and have potent inhibitory effects on tumor growth and vascularization.cancer ͉ chemotherapy ͉ hypoxia ͉ xenograft

show abstract

Section: Discussionmentioning

confidence: 99%

RETRACTED: Acriflavine inhibits HIF-1 dimerization, tumor growth, and vascularization

Lee¹,

Zhang²,

Qian³

et al. 2009

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

434

399

View full text Add to dashboard Cite

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“…In addition to changes in the primary structure of BdlA, our findings indicated changes in the tertiary/quaternary structure as truncated BdlA polypeptide chains oligomerized, probably via the PAS domains. It is unclear whether the truncated BdlA polypeptides are required to interact to enable dispersion, but the ability of PAS domains to oligomerize has been demonstrated previously to convert various input stimuli into signals that propagate to downstream components by modifying protein-protein interactions (22,23). To our knowledge, such extensive posttranslational modification involving cleavage combined with changes in the protein structure have not been described for bacterial proteins.…”

Section: Discussionmentioning

confidence: 97%

Dispersion by Pseudomonas aeruginosa requires an unusual posttranslational modification of BdlA

Petrova

Sauer

2012

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

124

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Dispersion enables biofilm bacteria to transit from the biofilm to the planktonic growth state and to spawn novel communities in new locales. Although the chemotaxis protein BdlA plays a role in the dispersion of Pseudomonas aeruginosa biofilms in response to environmental cues, little is known about regulation of BdlA activity or how BdlA modulates the dispersion response. Here, we demonstrate that BdlA in its native form is inactive and is activated upon nonprocessive proteolysis at a ClpP-protease-like cleavage site located between the Per Arnt Sim (PAS) sensory domains PASa and PASb. Activation of BdlA to enable biofilm dispersion requires phosphorylation at tyrosine-238 as a signal, elevated c-di-GMP levels, the chaperone ClpD, and the protease ClpP. The resulting truncated BdlA polypeptide chains directly interact and are required for P. aeruginosa biofilms to disperse. Our results provide a basis for understanding the mechanism of biofilm dispersion that may be applicable to a large number of biofilm-forming pathogenic species. Insights into the mechanism of BdlA function have implications for the control of biofilm-related infections.

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“…The structure was determined using the coordinates of our previously solved HIF-1a PasB domain (data not shown), which crystallizes as a homodimer, along with the published structure of ARNT PasB domain. 41 The HIF-1a/ARNT complex and 155 water molecules were refined using CNS 42 to R/R free values of 21.5/24.8, with rmsd bond and angle deviations of 0.004 Å and 0.80 , respectively.…”

Section: Methodsmentioning

confidence: 99%

Identification of Cys255 in HIF‐1α as a novel site for development of covalent inhibitors of HIF‐1α/ARNT PasB domain protein–protein interaction

et al. 2012

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The heterodimer HIF-1a (hypoxia inducible factor)/HIF-b (also known as ARNT-aryl hydrocarbon nuclear translocator) is a key mediator of cellular response to hypoxia. The interaction between these monomer units can be modified by the action of small molecules in the binding interface between their C-terminal heterodimerization (PasB) domains. Taking advantage of the presence of several cysteine residues located in the allosteric cavity of HIF-1a PasB domain, we applied a cysteine-based reactomics ''hotspot identification'' strategy to locate regions of HIF1a PasB domain critical for its interaction with ARNT. COMPOUND 5 was identified using a mass spectrometry-based primary screening strategy and was shown to react specifically with Cys255 of the HIF-1a PasB domain. Biophysical characterization of the interaction between PasB domains of HIF-1a and ARNT revealed that covalent binding of COMPOUND 5 to Cys255 reduced binding affinity between HIF-1a and ARNT PasB domains approximately 10-fold. Detailed NMR structural analysis of HIF-1a-PasB-COMPOUND 5 conjugate showed significant local conformation changes in the HIF-1a associated with key residues involved in the HIF-1a/ARNT PasB domain interaction as revealed by the crystal structure of the HIF-1a/ARNT PasB heterodimer. Our screening strategy could be applied to other targets to identify pockets surrounding reactive cysteines suitable for development of small molecule modulators of protein function.

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Structural Basis of ARNT PAS-B Dimerization: Use of a Common Beta-sheet Interface for Hetero- and Homodimerization

Cited by 125 publications

References 48 publications

RETRACTED: Acriflavine inhibits HIF-1 dimerization, tumor growth, and vascularization

RETRACTED: Acriflavine inhibits HIF-1 dimerization, tumor growth, and vascularization

Dispersion by Pseudomonas aeruginosa requires an unusual posttranslational modification of BdlA

Identification of Cys255 in HIF‐1α as a novel site for development of covalent inhibitors of HIF‐1α/ARNT PasB domain protein–protein interaction

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