SASDF27: Brain and Muscle ARNT-Like 1 from Mus musculus (D530-L625), monom...

基本情報

• 登録情報: データベース: SASBDB / ID: SASDF27

試料

Brain and Muscle ARNT-Like 1 from Mus musculus (D530-L625), monomer, trans-conformation locking mutation P624A

• Aryl hydrocarbon receptor nuclear translocator-like protein 1 (protein), BMAL1 P624A, Mus musculus

• 機能・相同性: Isoform 2 of Basic helix-loop-helix ARNT-like protein 1
• 生物種: Mus musculus (ハツカネズミ)
• 引用: ジャーナル: J Biol Chem / 年: 2019; タイトル: Structural and mechanistic insights into the interaction of the circadian transcription factor BMAL1 with the KIX domain of the CREB-binding protein.; 著者: Archit Garg / Roberto Orru / Weixiang Ye / Ute Distler / Jeremy E Chojnacki / Maja Köhn / Stefan Tenzer / Carsten Sönnichsen / Eva Wolf / ; 要旨: The mammalian CLOCK:BMAL1 transcription factor complex and its coactivators CREB-binding protein (CBP)/p300 and mixed-lineage leukemia 1 (MLL1) critically regulate circadian transcription and chromatin modification. Circadian oscillations are regulated by interactions of BMAL1's C-terminal transactivation domain (TAD) with the KIX domain of CBP/p300 (activating) and with the clock protein CRY1 (repressing) as well as by the BMAL1 G-region preceding the TAD. Circadian acetylation of Lys within the G-region enhances repressive BMAL1-TAD-CRY1 interactions. Here, we characterized the interaction of the CBP-KIX domain with BMAL1 proteins, including the BMAL1-TAD, parts of the G-region, and Lys Tethering the small compound 1-10 in the MLL-binding pocket of the CBP-KIX domain weakened BMAL1 binding, and MLL1-bound KIX did not form a ternary complex with BMAL1, indicating that the MLL-binding pocket is important for KIX-BMAL1 interactions. Small-angle X-ray scattering (SAXS) models of BMAL1 and BMAL1:KIX complexes revealed that the N-terminal BMAL1 G-region including Lys forms elongated extensions emerging from the bulkier BMAL1-TAD:KIX core complex. Fitting high-resolution KIX domain structures into the SAXS-derived envelopes suggested that the G-region emerges near the MLL-binding pocket, further supporting a role of this pocket in BMAL1 binding. Additionally, mutations in the second CREB-pKID/c-Myb-binding pocket of the KIX domain moderately impacted BMAL1 binding. The BMAL1(K537Q) mutation mimicking Lys acetylation, however, did not affect the KIX-binding affinity, in contrast to its enhancing effect on CRY1 binding. Our results significantly advance the mechanistic understanding of the protein interaction networks controlling CLOCK:BMAL1- and CBP-dependent gene regulation in the mammalian circadian clock.

登録者

• Archit Garg (Institute of Molecular Biology Mainz, Germany)

モデル

• モデル #3049: ; タイプ: dummy / ダミー原子の半径: 2.50 A / カイ2乗値: 1.139 / P-value: 0.983057; Omokage検索でこの集合体の類似形状データを探す (詳細)

試料

• 試料: 名称: Brain and Muscle ARNT-Like 1 from Mus musculus (D530-L625), monomer, trans-conformation locking mutation P624A; 試料濃度: 3.27 mg/ml
• バッファ: 名称: 25 mM Hepes, 150 NaCl, 1 mM DTT, 5% Glycerol / pH: 7.2

要素 #1676

名称: BMAL1 P624A / タイプ: protein
記述: Aryl hydrocarbon receptor nuclear translocator-like protein 1
分子量: 9.853 / 分子数: 1 / 由来: Mus musculus / 参照: UniProt: Q9WTL8-2
配列:

GPDASSPGGK KILNGGTPDI PSTGLLPGQA QETPGYPYSD SSSILGENPH IGIDMIDNDQ GSSSPSNDEA AMAVIMSLLE ADAGLGGPVD FSDLPWAL

実験情報

• ビーム: 設備名称: PETRA III EMBL P12 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron / 波長: 0.124 Å / スペクトロメータ・検出器間距離: 3 mm
• 検出器: 名称: Pilatus 2M

スキャン

タイトル: Brain and Muscle ARNT-Like 1 from Mus musculus (D530-L625), monomer, trans-conformation locking mutation P624A
測定日: 2017年5月27日 / 保管温度: 10.3 °C / セル温度: 10.1 °C / 照射時間: 0.045 sec. / フレーム数: 28 / 単位: 1/nm /

	Min	Max
Q	0.0611	4.4195

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 1019 /

	Min	Max
Q	0.121631	2.92089
P(R) point	1	1019
R	0	11

結果

Experimental MW: 13.45 kDa / カーブのタイプ: single_conc

	P(R)	Guinier	Guinier error
前方散乱 I0	2592	2554.73	7.06
慣性半径, Rg	2.957 nm	2.77 nm	0.09

	Min	Max
D	-	11
Guinier point	9	149