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基本情報
登録情報 | データベース: EMDB / ID: EMD-30376 | |||||||||
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タイトル | Cryo-EM structure of anEscherichia coli RNAP-promoter open complex (RPo) | |||||||||
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![]() | RNA polymerase / Transcription initiation complex / TRANSCRIPTION / TRANSCRIPTION-DNA complex | |||||||||
機能・相同性 | ![]() sigma factor antagonist complex / RNA polymerase complex / submerged biofilm formation / cellular response to cell envelope stress / cytosolic DNA-directed RNA polymerase complex / regulation of DNA-templated transcription initiation / sigma factor activity / bacterial-type flagellum assembly / bacterial-type flagellum-dependent cell motility / nitrate assimilation ...sigma factor antagonist complex / RNA polymerase complex / submerged biofilm formation / cellular response to cell envelope stress / cytosolic DNA-directed RNA polymerase complex / regulation of DNA-templated transcription initiation / sigma factor activity / bacterial-type flagellum assembly / bacterial-type flagellum-dependent cell motility / nitrate assimilation / DNA-directed RNA polymerase complex / transcription elongation factor complex / regulation of DNA-templated transcription elongation / transcription antitermination / DNA-templated transcription initiation / cell motility / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / response to heat / protein-containing complex assembly / intracellular iron ion homeostasis / protein dimerization activity / response to antibiotic / negative regulation of DNA-templated transcription / DNA-templated transcription / magnesium ion binding / DNA binding / zinc ion binding / membrane / metal ion binding / cytoplasm / cytosol 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / 解像度: 3.58 Å | |||||||||
![]() | Lin W / Feng Y | |||||||||
![]() | ![]() タイトル: Structural basis for transcription inhibition by E. coli SspA. 著者: Fulin Wang / Jing Shi / Dingwei He / Bei Tong / Chao Zhang / Aijia Wen / Yu Zhang / Yu Feng / Wei Lin / ![]() 要旨: Stringent starvation protein A (SspA) is an RNA polymerase (RNAP)-associated protein involved in nucleotide metabolism, acid tolerance and virulence of bacteria. Despite extensive biochemical and ...Stringent starvation protein A (SspA) is an RNA polymerase (RNAP)-associated protein involved in nucleotide metabolism, acid tolerance and virulence of bacteria. Despite extensive biochemical and genetic analyses, the precise regulatory role of SspA in transcription is still unknown, in part, because of a lack of structural information for bacterial RNAP in complex with SspA. Here, we report a 3.68 Å cryo-EM structure of an Escherichia coli RNAP-promoter open complex (RPo) with SspA. Unexpectedly, the structure reveals that SspA binds to the E. coli σ70-RNAP holoenzyme as a homodimer, interacting with σ70 region 4 and the zinc binding domain of EcoRNAP β' subunit simultaneously. Results from fluorescent polarization assays indicate the specific interactions between SspA and σ70 region 4 confer its σ selectivity, thereby avoiding its interactions with σs or other alternative σ factors. In addition, results from in vitro transcription assays verify that SspA inhibits transcription probably through suppressing promoter escape. Together, the results here provide a foundation for understanding the unique physiological function of SspA in transcription regulation in bacteria. | |||||||||
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構造の表示
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構造ビューア | EMマップ: ![]() ![]() ![]() |
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マップデータ | ![]() | 28.5 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 19.3 KB 19.3 KB | 表示 表示 | ![]() |
画像 | ![]() | 123.6 KB | ||
Filedesc metadata | ![]() | 8 KB | ||
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-検証レポート
文書・要旨 | ![]() | 576.5 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 576.1 KB | 表示 | |
XML形式データ | ![]() | 5.6 KB | 表示 | |
CIF形式データ | ![]() | 6.4 KB | 表示 | |
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「今月の分子」の関連する項目 |
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マップ
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ボクセルのサイズ | X=Y=Z: 1.307 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
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試料の構成要素
+全体 : Escherichia coli RNAP-promoter open complex (RPo)
+超分子 #1: Escherichia coli RNAP-promoter open complex (RPo)
+分子 #1: DNA (63-MER)
+分子 #7: DNA (63-MER)
+分子 #2: DNA-directed RNA polymerase subunit alpha
+分子 #3: DNA-directed RNA polymerase subunit beta
+分子 #4: DNA-directed RNA polymerase subunit beta'
+分子 #5: DNA-directed RNA polymerase subunit omega
+分子 #6: RNA polymerase sigma factor RpoD
+分子 #8: MAGNESIUM ION
+分子 #9: ZINC ION
-実験情報
-構造解析
![]() | 単粒子再構成法 |
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試料の集合状態 | particle |
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試料調製
緩衝液 | pH: 7.9 |
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電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: COUNTING / 平均電子線量: 59.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |