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基本情報
登録情報 | ![]() | |||||||||
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タイトル | M. tuberculosis ribosomal RNA methyltransferase TlyA bound to M. smegmatis 50S ribosomal subunit | |||||||||
![]() | Mtb TlyA bound to Msg 50S ribosomal subunit. | |||||||||
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![]() | Ribosome / Methyltransferase / rRNA modification / 50S / TRANSFERASE | |||||||||
機能・相同性 | ![]() 23S rRNA (cytidine1920-2'-O)-methyltransferase / 16S rRNA (cytidine1409-2'-O)-methyltransferase / methyltransferase activity / methylation / cytosolic large ribosomal subunit / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation ...23S rRNA (cytidine1920-2'-O)-methyltransferase / 16S rRNA (cytidine1409-2'-O)-methyltransferase / methyltransferase activity / methylation / cytosolic large ribosomal subunit / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / mRNA binding / RNA binding / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.05 Å | |||||||||
![]() | Laughlin ZT / Dunham CM | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: 50S subunit recognition and modification by the ribosomal RNA methyltransferase TlyA. 著者: Zane T Laughlin / Suparno Nandi / Debayan Dey / Natalia Zelinskaya / Marta A Witek / Pooja Srinivas / Ha An Nguyen / Emily G Kuiper / Lindsay R Comstock / Christine M Dunham / Graeme L Conn / ![]() 要旨: Changes in bacterial ribosomal RNA (rRNA) methylation status can alter the activity of diverse groups of ribosome-targeting antibiotics. These modifications are typically incorporated by a single ...Changes in bacterial ribosomal RNA (rRNA) methylation status can alter the activity of diverse groups of ribosome-targeting antibiotics. These modifications are typically incorporated by a single methyltransferase that acts on one nucleotide target and rRNA methylation directly prevents drug binding, thereby conferring drug resistance. Loss of intrinsic methylation can also result in antibiotic resistance. For example, Mycobacterium tuberculosis becomes sensitized to tuberactinomycin antibiotics, such as capreomycin and viomycin, due to the action of the intrinsic methyltransferase TlyA. TlyA is unique among antibiotic resistance-associated methyltransferases as it has dual 16S and 23S rRNA substrate specificity and can incorporate cytidine-2′-O-methylations within two structurally distinct contexts. Here, we report the structure of a mycobacterial 50S subunit-TlyA complex trapped in a postcatalytic state with a S-adenosyl-L-methionine analog using single-particle cryogenic electron microscopy. Together with complementary functional analyses, this structure reveals critical roles in 23S rRNA substrate recognition for conserved residues across an interaction surface that spans both TlyA domains. These interactions position the TlyA active site over the target nucleotide C2144, which is flipped from 23S Helix 69 in a process stabilized by stacking of TlyA residue Phe157 on the adjacent A2143. Base flipping may thus be a common strategy among rRNA methyltransferase enzymes, even in cases where the target site is accessible without such structural reorganization. Finally, functional studies with 30S subunit suggest that the same TlyA interaction surface is employed to recognize this second substrate, but with distinct dependencies on essential conserved residues. #1: ![]() タイトル: 50S subunit recognition and modification by the Mycobacterium tuberculosis ribosomal RNA methyltransferase TlyA 著者: Laughlin ZT / Nandi S / Dey D / Zelinskaya N / Witek MA / Srinivas P / Nguyen HA / Kuiper EG / Comstock LR / Dunham CM / Conn GL | |||||||||
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マップデータ | ![]() | 78.6 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 62.4 KB 62.4 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 9.9 KB | 表示 | ![]() |
画像 | ![]() | 82.3 KB | ||
Filedesc metadata | ![]() | 12.7 KB | ||
その他 | ![]() ![]() | 77.3 MB 78.3 MB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 7s0sMC M: このマップから作成された原子モデル C: 同じ文献を引用 ( |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||
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注釈 | Mtb TlyA bound to Msg 50S ribosomal subunit. | ||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.069 Å | ||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-追加マップ: Mtb TlyA bound to H69 and proximal rRNA...
ファイル | emd_24792_additional_1.map | ||||||||||||
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注釈 | Mtb TlyA bound to H69 and proximal rRNA without the rest of Msg 50S. Created from multi body refinement and used in model building and refinement. | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Msg 50S alone without H69 or Mtb TlyA....
ファイル | emd_24792_additional_2.map | ||||||||||||
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注釈 | Msg 50S alone without H69 or Mtb TlyA. Created from multi body refinement. | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
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試料の構成要素
+全体 : Mtb TlyA bound to Msg 50S ribosomal subunit modified at C2144 wit...
+超分子 #1: Mtb TlyA bound to Msg 50S ribosomal subunit modified at C2144 wit...
+超分子 #2: 50S
+超分子 #3: TlyA
+分子 #1: ribosomal protein bL37
+分子 #2: 16S/23S rRNA (Cytidine-2'-O)-methyltransferase TlyA
+分子 #4: 50S ribosomal protein L2
+分子 #5: 50S ribosomal protein L3
+分子 #6: 50S ribosomal protein L4
+分子 #7: 50S ribosomal protein L5
+分子 #8: 50S ribosomal protein L6
+分子 #9: 50S ribosomal protein L9
+分子 #10: 50S ribosomal protein L10
+分子 #11: 50S ribosomal protein L11
+分子 #12: 50S ribosomal protein L13
+分子 #13: 50S ribosomal protein L14
+分子 #14: 50S ribosomal protein L15
+分子 #15: 50S ribosomal protein L16
+分子 #16: 50S ribosomal protein L17
+分子 #17: 50S ribosomal protein L18
+分子 #18: 50S ribosomal protein L19
+分子 #19: 50S ribosomal protein L20
+分子 #20: 50S ribosomal protein L21
+分子 #21: 50S ribosomal protein L22
+分子 #22: 50S ribosomal protein L23
+分子 #23: 50S ribosomal protein L24
+分子 #24: 50S ribosomal protein L25
+分子 #25: 50S ribosomal protein L27
+分子 #26: 50S ribosomal protein L28
+分子 #27: 50S ribosomal protein L29
+分子 #28: 50S ribosomal protein L30
+分子 #29: 50S ribosomal protein L32
+分子 #30: 50S ribosomal protein L33
+分子 #31: 50S ribosomal protein L34
+分子 #32: 50S ribosomal protein L35
+分子 #33: 50S ribosomal protein L36
+分子 #34: 50S ribosomal protein L31
+分子 #3: 23S rRNA
+分子 #35: 5S rRNA
+分子 #36: MAGNESIUM ION
+分子 #37: ZINC ION
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
緩衝液 | pH: 7 構成要素:
詳細: Both components dialyzed into this buffer | ||||||||||||
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グリッド | モデル: Quantifoil R1.2/1.3 / 材質: COPPER / 前処理 - タイプ: GLOW DISCHARGE | ||||||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 90 % / チャンバー内温度: 298 K / 装置: GATAN CRYOPLUNGE 3 / 詳細: 3.0 or 3.3 second blot time allowed for sample. | ||||||||||||
詳細 | 0.5 micromolar 50S, 5 micromolar TlyA, 10 micromolar NM6 |
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電子顕微鏡法
顕微鏡 | TFS KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K3 (6k x 4k) / デジタル化 - サイズ - 横: 5760 pixel / デジタル化 - サイズ - 縦: 4092 pixel / 撮影したグリッド数: 2 / 実像数: 3364 / 平均露光時間: 2.0 sec. / 平均電子線量: 50.79 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 / Cs: 2.7 mm / 最大 デフォーカス(公称値): 2.2 µm / 最小 デフォーカス(公称値): 0.8 µm / 倍率(公称値): 81000 |
試料ステージ | ホルダー冷却材: NITROGEN |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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画像解析
-原子モデル構築 1
初期モデル |
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得られたモデル | ![]() PDB-7s0s: |