Heme-Binding Characteristics of the Isolated PAS-A Domain of Mouse Per2, a Transcriptional Regulatory Factor Associated with Circadian Rhythms

Kitanishi, Kenichi; Igarashi, Jun–ichi; Hayasaka, Koya; Hikage, Naoki; Saiful, Islam S. M.; Yamauchi, Seigo; Uchida, Takeshi; Ishimori, Koichiro; Shimizu, Tôru

doi:10.1021/bi7023892

Cited by 56 publications

(64 citation statements)

References 40 publications

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“…Although mPER proteins do not have known sensory functions, mPER2 has been reported to bind heme as a cofactor in its PAS domains and in its C-terminal region (13,14). Our UV/ VIS spectroscopic analyses (Fig.…”

Section: Discussionmentioning

confidence: 86%

“…S7D). Cys270, another potential heme ligand of mPER2 (13), is conserved in mPER1 and mPER3 (Cys299 mPER1 , Cys210 mPER3 ). Our mPER crystal structures will guide the design of His and Cys mutants to evaluate heme binding in vivo and in vitro.…”

Section: Discussionmentioning

confidence: 99%

“…These interactions regulate the stability and cellular localization of the mPERs and modulate the activity of the mBMAL1/mCLOCK transcription factor complex. Additionally, the PAS domains of NPAS2, mCLOCK, and mPER2 have been reported to bind heme (13)(14)(15)(16). In the circadian clock, mPER1 and mPER2 proteins are found in large protein complexes, likely establishing multiple interactions via their PAS domains, the central CKIε∕δ binding domain and the C-terminal mCRY binding region (17)(18)(19).…”

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confidence: 99%

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Unwinding the differences of the mammalian PERIOD clock proteins from crystal structure to cellular function

Kucera

Schmalen

Hennig

et al. 2012

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

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The three PERIOD homologues mPER1, mPER2, and mPER3 constitute central components of the mammalian circadian clock. They contain two PAS (PER-ARNT-SIM) domains (PAS-A and PAS-B), which mediate homo-and heterodimeric mPER-mPER interactions as well as interactions with transcription factors and kinases. Here we present crystal structures of PAS domain fragments of mPER1 and mPER3 and compare them with the previously reported mPER2 structure. The structures reveal homodimers, which are mediated by interactions of the PAS-B β-sheet surface including a highly conserved tryptophan (Trp448 mPER1 , Trp419 mPER2 , Trp359 mPER3 ). mPER1 homodimers are additionally stabilized by interactions between the PAS-A domains and mPER3 homodimers by an N-terminal region including a predicted helix-loop-helix motive. We have verified the existence of these homodimer interfaces in solution and inside cells using analytical gel filtration and luciferase complementation assays and quantified their contributions to homodimer stability by analytical ultracentrifugation. We also show by fluorescence recovery after photobleaching analyses that destabilization of the PAS-B/tryptophan dimer interface leads to a faster mobility of mPER2 containing complexes in human U2OS cells. Our study reveals structural and quantitative differences between the homodimeric interactions of the three mouse PERIOD homologues, which are likely to contribute to their distinct clock functions.circadian clock | PAS domains | PERIOD proteins | protein interactions

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Section: Discussionmentioning

confidence: 86%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Unwinding the differences of the mammalian PERIOD clock proteins from crystal structure to cellular function

Kucera

Schmalen

Hennig

et al. 2012

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

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“…These redox-and ligand-dependent changes could then trigger alterations in circadian patterns and, if not reversed, result in circadian and/or metabolic disorders. The effects of heme, redox, and intrinsic and extrinsic (CO and NO) ligands on RevErb would likely be complemented by those on other heme proteins with key roles in the circadian cycle, such as NPAS2 and Per2 (82,83).…”

Section: Discussionmentioning

confidence: 99%

Thiol-disulfide Redox Dependence of Heme Binding and Heme Ligand Switching in Nuclear Hormone Receptor Rev-erbβ

Gupta

Ragsdale

2011

Journal of Biological Chemistry

118

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Rev-erb␤ is a heme-binding nuclear hormone receptor that represses a broad spectrum of target genes involved in regulating metabolism, the circadian cycle, and proinflammatory responses. Here, we demonstrate that a thiol-disulfide redox switch controls the interaction between heme and the ligandbinding domain of Rev-erb␤. The reduced dithiol state of Rev-erb␤ binds heme 5-fold more tightly than the oxidized disulfide state. By means of site-directed mutagenesis and by UV-visible and EPR spectroscopy, we also show that the ferric heme of reduced (dithiol) Rev-erb␤ can undergo a redox-triggered switch from imidazole/thiol ligation (via His-568 and Cys-384, based on a prior crystal structure) to His/neutral residue ligation upon oxidation to the disulfide form. On the other hand, we find that change in the redox state of iron has no effect on heme binding to the ligand-binding domain of the protein. The low dissociation constant for the complex between Fe 3؉ -or Fe 2؉ -heme and the reduced dithiol state of the protein (K d ‫؍‬ ϳ20 nM) is in the range of the intracellular free heme concentration. We also determined that the Fe 2؉ -heme bound to the ligand-binding domain of Rev-erb␤ has high affinity for CO (K d ‫؍‬ 60 nM), which replaces one of the internal ligands when bound. We suggest that this thiol-disulfide redox switch is one mechanism by which oxidative stress is linked to circadian and/or metabolic imbalance. Heme dissociation from Rev-erb␤ has been shown to derepress the expression of target genes in response to changes in intracellular redox conditions. We propose that oxidative stress leads to oxidation of cysteine(s), thus releasing heme from Rev-erb␤ and altering its transcriptional activity.Nuclear receptors regulate gene expression in response to small molecules, controlling key eukaryotic events such as development, differentiation, reproduction, and metabolic homeostasis (1). All nuclear receptors possess a highly conserved N-terminal DNA-binding domain and a less conserved C-terminal ligand-binding domain (LBD), 2 which recognizes small molecules (2). The LBD in nuclear receptors is typically coupled to transcriptional activation through an activation function-2 domain, which promotes recruitment of a coactivator (1). Rev-erb␤ (also known as RVR and BD73) and Reverb␣ are unique members of the superfamily of nuclear receptors because they lack an activation function-2 domain (3, 4).Rev-erbs are transcriptional repressors that are present in the nucleus and bind as a monomer to the Rev responsive element or as a dimer to a Rev-RE direct repeat, RevDR-2 (5, 6). Rev-erbs had been considered to be an orphan nuclear receptor until the Drosophila ortholog of human Rev-erb (E75) (7) and then Rev-erb ␣ (8, 9) and Rev-erb␤ (8) were recently demonstrated to bind heme at their LBDs. Heme binding promotes recruitment of nuclear corepressor and histone deacetylase (HDAC) (8, 10, 11). Rev-erb␣ recruits HDAC3, whereas Rev-erb␤ recruits HDAC1 (3,8,9). In turn, the Reverb-nuclear corepressor-HDAC complex facilitate...

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“…With respect to Per2, this molecule contains three heme binding regions: two PAS domains and a C-terminal HRM containing a CP motif. The binding of heme to the Per2 PAS domains appears to regulate DNA binding in response to carbon monoxide (Kitanishi et al 2008;Hayasaka et al 2011). Meanwhile, heme binding to the C-terminal HRM stimulates the degradation of Per2 .…”

Section: Circadian and Metabolic Regulatorsmentioning

confidence: 99%

Wearing Red for Signaling: The Heme-Bach Axis in Heme Metabolism, Oxidative Stress Response and Iron Immunology

Igarashi

Watanabe-Matsui

2014

Tohoku J. Exp. Med.

102

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The connection between gene regulation and metabolism is an old issue that warrants revisiting in order to understand both normal as well as pathogenic processes in higher eukaryotes. Metabolites affect the gene expression by either binding to transcription factors or serving as donors for post-translational modification, such as that involving acetylation and methylation. The focus of this review is heme, a prosthetic group of proteins that includes hemoglobin and cytochromes. Heme has been shown to bind to several transcription factors, including Bach1 and Bach2, in higher eukaryotes. Heme inhibits the transcriptional repressor activity of Bach1, resulting in the derepression of its target genes, such as globin in erythroid cells and heme oxygenase-1 in diverse cell types. Since Bach2 is important for class switch recombination and somatic hypermutation of immunoglobulin genes as well as regulatory and effector T cell differentiation and the macrophage function, the heme-Bach2 axis may regulate the immune response as a signaling cascade. We discuss future issues regarding the topic of the iron/heme-gene regulation network based on current understanding of the heme-Bach axis, including the concept of "iron immunology" as the synthesis of the iron metabolism and the immune response.

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Heme-Binding Characteristics of the Isolated PAS-A Domain of Mouse Per2, a Transcriptional Regulatory Factor Associated with Circadian Rhythms

Cited by 56 publications

References 40 publications

Unwinding the differences of the mammalian PERIOD clock proteins from crystal structure to cellular function

Unwinding the differences of the mammalian PERIOD clock proteins from crystal structure to cellular function

Thiol-disulfide Redox Dependence of Heme Binding and Heme Ligand Switching in Nuclear Hormone Receptor Rev-erbβ

Wearing Red for Signaling: The Heme-Bach Axis in Heme Metabolism, Oxidative Stress Response and Iron Immunology

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