National Natural Science Foundation of China (NSFC)
31770068, 32070040
中国
引用
ジャーナル: Nucleic Acids Res / 年: 2022 タイトル: Structural basis of Streptomyces transcription activation by zinc uptake regulator. 著者: Xu Yang / Yiqun Wang / Guiyang Liu / Zixin Deng / Shuangjun Lin / Jianting Zheng / 要旨: Streptomyces coelicolor (Sc) is a model organism of actinobacteria to study morphological differentiation and production of bioactive metabolites. Sc zinc uptake regulator (Zur) affects both ...Streptomyces coelicolor (Sc) is a model organism of actinobacteria to study morphological differentiation and production of bioactive metabolites. Sc zinc uptake regulator (Zur) affects both processes by controlling zinc homeostasis. It activates transcription by binding to palindromic Zur-box sequences upstream of -35 elements. Here we deciphered the molecular mechanism by which ScZur interacts with promoter DNA and Sc RNA polymerase (RNAP) by cryo-EM structures and biochemical assays. The ScZur-DNA structures reveal a sequential and cooperative binding of three ScZur dimers surrounding a Zur-box spaced 8 nt upstream from a -35 element. The ScRNAPσHrdB-Zur-DNA structures define protein-protein and protein-DNA interactions involved in the principal housekeeping σHrdB-dependent transcription initiation from a noncanonical promoter with a -10 element lacking the critical adenine residue at position -11 and a TTGCCC -35 element deviating from the canonical TTGACA motif. ScZur interacts with the C-terminal domain of ScRNAP α subunit (αCTD) in a complex structure trapped in an active conformation. Key ScZur-αCTD interfacial residues accounting for ScZur-dependent transcription activation were confirmed by mutational studies. Together, our structural and biochemical results provide a comprehensive model for transcription activation of Zur family regulators.
全体 : Local refinement map of Zur regions in Streptomyces coelicolor RN...
全体
名称: Local refinement map of Zur regions in Streptomyces coelicolor RNAP-promoter open complex with three Zur dimers
要素
複合体: Local refinement map of Zur regions in Streptomyces coelicolor RNAP-promoter open complex with three Zur dimers
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超分子 #1: Local refinement map of Zur regions in Streptomyces coelicolor RN...
超分子
名称: Local refinement map of Zur regions in Streptomyces coelicolor RNAP-promoter open complex with three Zur dimers タイプ: complex / キメラ: Yes / ID: 1 / 親要素: 0 / 含まれる分子: #1-#9
由来(天然)
生物種: Streptomyces coelicolor A3(2) (バクテリア)
組換発現
生物種: Escherichia coli BL21(DE3) (大腸菌)
分子量
実験値: 560 KDa
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実験情報
-
構造解析
手法
クライオ電子顕微鏡法
解析
単粒子再構成法
試料の集合状態
particle
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試料調製
濃度
0.8 mg/mL
緩衝液
pH: 8 構成要素:
濃度
式
名称
20.0 mM
HEPES
HEPES
100.0 mM
KCl
KCl
2.0 mM
DTT
DTT
5.0 mM
MgCl2
MgCl2
0.02 mM
ZnSO4
ZnSO4
凍結
凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 289 K / 装置: FEI VITROBOT MARK IV
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
撮影
フィルム・検出器のモデル: GATAN K2 QUANTUM (4k x 4k) 平均電子線量: 1.25 e/Å2