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基本情報
登録情報 | データベース: EMDB / ID: EMD-0329 | |||||||||
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タイトル | Rea1 AAA2L-H2alpha deletion mutant in AMPPNP State | |||||||||
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![]() | Rea1 / Mdn1 / Midasin / AAA+ protein / ribosome maturation / molecular machine / MOTOR PROTEIN | |||||||||
機能・相同性 | ![]() protein-RNA complex remodeling / regulation of ribosomal subunit export from nucleus / preribosome, large subunit precursor / ribosomal large subunit export from nucleus / rRNA processing / nucleolus / ATP hydrolysis activity / mitochondrion / nucleoplasm / ATP binding / nucleus 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: 7.8 Å | |||||||||
![]() | Sosnowski P / Urnavicius L | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: The CryoEM structure of the ribosome maturation factor Rea1. 著者: Piotr Sosnowski / Linas Urnavicius / Andreas Boland / Robert Fagiewicz / Johan Busselez / Gabor Papai / Helgo Schmidt / ![]() ![]() 要旨: The biogenesis of 60S ribosomal subunits is initiated in the nucleus where rRNAs and proteins form pre-60S particles. These pre-60S particles mature by transiently interacting with various assembly ...The biogenesis of 60S ribosomal subunits is initiated in the nucleus where rRNAs and proteins form pre-60S particles. These pre-60S particles mature by transiently interacting with various assembly factors. The ~5000 amino-acid AAA+ ATPase Rea1 (or Midasin) generates force to mechanically remove assembly factors from pre-60S particles, which promotes their export to the cytosol. Here we present three Rea1 cryoEM structures. We visualise the Rea1 engine, a hexameric ring of AAA+ domains, and identify an α-helical bundle of AAA2 as a major ATPase activity regulator. The α-helical bundle interferes with nucleotide-induced conformational changes that create a docking site for the substrate binding MIDAS domain on the AAA +ring. Furthermore, we reveal the architecture of the Rea1 linker, which is involved in force generation and extends from the AAA+ ring. The data presented here provide insights into the mechanism of one of the most complex ribosome maturation factors. | |||||||||
履歴 |
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構造の表示
ムービー |
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構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
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マップデータ | ![]() | 5.5 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 20.5 KB 20.5 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 9.2 KB | 表示 | ![]() |
画像 | ![]() | 30.4 KB | ||
Filedesc metadata | ![]() | 9.1 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
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リンク
EMDBのページ | ![]() ![]() |
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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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ボクセルのサイズ | X=Y=Z: 1.635 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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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 : Rea1 (MIDASIN) AAA2L-H2alpha deletion mutant in AMPPNP State - el...
全体 | 名称: Rea1 (MIDASIN) AAA2L-H2alpha deletion mutant in AMPPNP State - electron density map |
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要素 |
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-超分子 #1: Rea1 (MIDASIN) AAA2L-H2alpha deletion mutant in AMPPNP State - el...
超分子 | 名称: Rea1 (MIDASIN) AAA2L-H2alpha deletion mutant in AMPPNP State - electron density map タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: all |
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由来(天然) | 生物種: ![]() ![]() |
-分子 #1: Midasin,Midasin,Midasin,Midasin,Midasin,Midasin,Midasin
分子 | 名称: Midasin,Midasin,Midasin,Midasin,Midasin,Midasin,Midasin タイプ: protein_or_peptide / ID: 1 / コピー数: 1 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 546.53825 KDa |
組換発現 | 生物種: ![]() ![]() |
配列 | 文字列: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) ...文字列: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)PSVPECFTI E KKSSYFII EPQDLSTKVA SICGVIVPKV HTIHDKVFYP LTFVPTHKTV SSLRQLGRKI QNSTPIMLIG KAGSGKTFLI NE LSKYMGC HDSIVKIHLG EQTDAKLLIG TYTSGDKPGT FEWRAGVLAT AVKEGRWVLI EDIDKAPTDV LSILLSLLEK REL TIPSRG ETVKAANGFQ LISTVRINED HQKDSSNKIY NLNMIGMRIW NVIELEEPSE EDLTHILAQK FPILTNLIPK LIDS YKNVK SIYMNTKFIS LNKGAHTRVV SVRDLIKLCE RLDILFKNNG INKPDQLIQS SVYDSIFSEA ADCFAGAIGE FKALE PIIQ AIGESLDIAS SRISLFLTQH VPTLENLDDS IKIGRAVLLK EKLNIQKKSM NSTLFAFTNH SLRLMEQISV CIQMTE PVL LVGETGTGKT TVVQQLAKML AKKLTVINVS QQTETGDLLG GYKPVNSKTV AVPIQENFET LFNATFSLKK NEKFHKM LH RCFNKNQWKN VVKLWNEAYK MAQSILKITN TENENENAKK KKRRLNTHEK KLLLDKWADF NDSVKKFEAQ SSSIENSF V FNFVEGSLVK TIRAGEWLLL DEVNLATADT LESISDLLTE PDSRSILLSE KGDAEPIKAH PDFRIFACMN PATDVGKRD LPMGIRSRFT EIYVHSPERD ITDLLSIIDK YIGKYSVSDE WVGNDIAELY LEAKKLSDNN TIVDGSNQKP HFSIRTLTRT LLYVTDIIH IYGLRRSLYD GFCMSFLTLL DQKSEAILKP VIEKFTLGRL KNVKSIMSQT PPSPGPDYVQ FKHYWMKKGP N TIQEQAHY IITPFVEKNM MNLVRATSGK RFPVLIQGPT SSGKTSMIKY LADITGHKFV RINNHEHTDL QEYLGTYVTD DT GKLSFKE GVLVEALRKG YWIVLDELNL APTDVLEALN RLLDDNRELF IPETQEVVHP HPDFLLFATQ NPPGIYGGRK ILS RAFRNR FLELHFDDIP QDELEIILRE RCQIAPSYAK KIVEVYRQLS IERSASRLFE QKNSFATLRD LFRWALRDAV GYEQ LAASG YMLLAERCRT PQEKVTVKKT LEKVMKVKLD MDQYYASLED KSLEAIGSVT WTKGMRRLSV LVSSCLKNKE PVLLV GETG CGKTTICQLL AQFMGRELIT LNAHQNTETG DILGAQRPVR NRSEIQYKLI KSLKTALNIA NDQDVDLKEL LQLYSK SDN KNIAEDVQLE IQKLRDSLNV LFEWSDGPLI QAMRTGNFFL LDEISLADDS VLERLNSVLE PERSLLLAEQ GSSDSLV TA SENFQFFATM NPGGDYGKKE LSPALRNRFT EIWVPSMEDF NDVNMIVSSR LLEDLKDLAN PIVKFSEWFG KKLGGGNA T SGVISLRDIL AWVEFINKVF PKIQNKSTAL IQGASMVFID ALGTNNTAYL AENENDLKSL RTECIIQLLK LCGDDLELQ QIETNEIIVT QDELQVGMFK IPRFPDAQSS SFNLTAPTTA SNLVRVVRAM QVHKPILLEG SPGVGKTSLI TALANITGNK LTRINLSEQ TDLVDLFGAD APGERSGEFL WHDAPFLRAM KKGEWVLLDE MNLASQSVLE GLNACLDHRG EAYIPELDIS F SCHPNFLV FAAQNPQYQG GGRKGLPKSF VNRFSVVFID MLTSDDLLLI AKHLYPSIEP DIIAKMIKLM STLEDQVCKR KL WGNSGSP WEFNLRDTLR WLKLLNQYSI CEDVDVFDFV DIIVKQRFRT ISDKNKAQLL IEDIFGKFST KENFFKLTED YVQ INNEVA LRNPHYRYPI TQNLFPLECN VAVYESVLKA INNNWPLVLV GPSNSGKTET IRFLASILGP RVDVFSMNSD IDSM DILGG YEQVDLTRQI SYITEELTNI VREIISMNMK LSPNATAIME GLNLLKYLLN NIVTPEKFQD FRNRFNRFFS HLEGH PLLK TMSMNIEKMT EIITKEASVK FEWFDGMLVK AVEKGHWLIL DNANLCSPSV LDRLNSLLEI DGSLLINECS QEDGQP RVL KPHPNFRLFL TMDPKYGELS RAMRNRGVEI YIDELHSRST AFDRLTLGFE LGENIDFVSI DDGIKKIKLN EPDMSIP LK HYVPSYLSRP CIFAQVHDIL LLSDEEPIEE SLAAVIPISH LGEVGKWANN VLNCTEYSEK KIAERLYVFI TFLTDMGV L EKINNLYKPA NLKFQKALGL HDKQLTEETV SLTLNEYVLP TVSKYSDKIK SPESLYLLSS LRLLLNSLNA LKLINEKST HGKIDELTYI ELSAAAFNGR HLKNIPRIPI FCILYNILTV MSENLKTESL FCGSNQYQYY WDLLVIVIAA LETAVTKDEA RLRVYKELI DSWIASVKSK SDIEITPFLN INLEFTDVLQ LSRGHSITLL WDIFRKNYPT TSNSWLAFEK LINLSEKFDK V RLLQFSES YNSIKDLMDV FRLLNDDVLN NKLSEFNLLL SKLEDGINEL ELISNKFLNK RKHYFADEFD NLIRYTFSVD TA ELIKELA PASSLATQKL TKLITNKYNY PPIFDVLWTE KNAKLTSFTS TIFSSQFLED VVRKSNNLKS FSGNQIKQSI SDA ELLLSS TIKCSPNLLK SQMEYYKNML LSWLRKVIDI HVGGDCLKLT LKELCSLIEE KTASETRVTF AEYIFPALDL AESS KSLEE LGEAWITFGT GLLLLFVPDS PYDPAIHDYV LYDLFLKTKT FSQNLMKSWR NVRKVISGDE EIFTEKLINT ISDDD APQS PRVYRTGMSI DSLFDEWMAF LSSTMSSRQI KELVSSYKCN SDQSDRRLEM LQQNSAHFLN RLESGYSKFA DLNDIL AGY IYSINFGFDL LKLQKSKDRA SFQISPLWSM DPINISCAEN VLSAYHELSR FFKKGDMEDT SIEKVLMYFL TLFKFHK RD TNLLEIFEAA LYTLYSRWSV RRFRQEQEEN EKSNMFKFND NSDDYEADFR KLFPDYEDTA LVTNEKDISS PENLDDIY F KLADTYISVF DKDHDANFSS ELKSGAIITT ILSEDLKNTR IEELKSGSLS AVINTLDAET QSFKNTEVFG NIDFYHDFS IPEFQKAGDI IETVLKSVLK LLKQWPEHAT LKELYRVSQE FLNYPIKTPL ARQLQKIEQI YTYLAEWEKY ASSEVSLNNT VKLITDLIV SWRKLELRTW KGLFNSEDAK TRKSIGKWWF YLYESIVISN FVSEKKETAP NATLLVSSLN LFFSKSTLGE F NARLDLVK AFYKHIQLIG LRSSKIAGLL HNTIKFYYQF KPLIDERITN GKKSLEKEID DIILLASWKD VNVDALKQSS RK SHNNLYK IVRKYRDLLN GDAKTIIEAG LLY(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)RNID TVASNMDSYL EKISSQEFPN FADLASDFYA EAERLRKETP NVYTKENKKR L AYLKTQKS KLLGDALKEL RRIGLKVNFR EDIQKVQSST TTILANIAPF NNEYLNSSDA FFFKILDLLP KLRSAASNPS DD IPVAAIE RGMALAQSLM FSLITVRHPL SEFTNDYCKI NGMMLDLEHF TCLKGDIVHS SLKANVDNVR LFEKWLPSLL DYA AQTLSV ISKYSATSEQ QKILLDAKST LSSFFVHFNS SRIFDSSFIE SYSRFELFIN ELLKKLENAK ETGNAFVFDI IIEW IKANK GGPIKKEQKR GPSVEDVEQA FRRTFTSIIL SFQKVIGDGI ESISETDDNW LSASFKKVMV NVKLLRSSVV SKNIE TALS LLKDFDFTTT ESIYVKSVIS FTLPVITRYY NAMTVVLERS RIYYTNTSRG MYILSTILHS LAKNGFCSPQ PPSEEV DDK NLQEGTGLGD GEGAQNNNKD VEQDEDLTED AQNENKEQQD KDERDDENED DAVEMEGDMA GELEDLSNGE ENDDEDT DS EEEELDEEID DLNEDDPNAI DDKMWDDKAS DNSKEKDTDQ NLDGKNQEED VQAAENDEQQ RDNKEGGDED PNAPEDGD E EIENDENAEE ENDVGEQEDE VKDEEGEDLE ANVPEIETLD LPEDMNLDSE HEESDEDVDM SDGMPDDLNK EEVGNEDEE VKQESGIESD NENDEPGPEE DAGETETALD EEEGAEEDVD MTNDEGKEDE ENGPEEQAMS DEEELKQDAA MEENKEKGGE QNTEGLDGV EEKADTEDID QEAAVQQDSG SKGAGADATD TQEQDDVGGS GTTQNTYEED QEDVTKNNEE SREEATAALK Q LGDSMKEY HRRRQDIKEA QTNGEEDENL EKNNERPDEF EHVEGANTET DTQALGSATQ DQLQTIDEDM AIDDDREEQE VD QKELVED ADDEKMDIDE EEMLSDIDAH DANNDVDSKK SGFIGKRKSE EDFENELSNE HFSADQEDDS EIQSLIENIE DNP PDASAS LTPERSLEES RELWHKSEIS TADLVSRLGE QLRLILEPTL ATKLKGDYKT GKRLNMKRII PYIASQFRKD KIWL RRTKP SKRQYQIMIA LDDSKSMSES KCVKLAFDSL CLVSKTLTQL EAGGLSIVKF GENIKEVHSF DQQFSNESGA RAFQW FGFQ ETKTDVKKLV AESTKIFERA RAMVHNDQWQ LEIVISDGIC EDHETIQKLV RRARENKIML VFVIIDGITS NESILD MSQ VNYIPDQYGN PQLKITKYLD TFPFEFYVVV HDISELPEML SLILRQYFTD LASS UniProtKB: Midasin, Midasin |
-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
濃度 | 1 mg/mL | ||||||||||||||||||
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緩衝液 | pH: 7.2 構成要素:
詳細: AMP-PNP was added 5 minute before the plunging | ||||||||||||||||||
グリッド | モデル: Quantifoil R2/2 / 材質: COPPER / メッシュ: 200 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY / 支持フィルム - Film thickness: 12 / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 時間: 30 sec. / 前処理 - 雰囲気: AIR | ||||||||||||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 90 % / チャンバー内温度: 280 K / 装置: FEI VITROBOT MARK III |
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電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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特殊光学系 | 位相板: VOLTA PHASE PLATE / 球面収差補正装置: Titan Krios Cs Corrector / エネルギーフィルター - 名称: GIF Quantum LS / エネルギーフィルター - スリット幅: 20 eV |
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 / デジタル化 - サイズ - 横: 3712 pixel / デジタル化 - サイズ - 縦: 3840 pixel / デジタル化 - 画像ごとのフレーム数: 2-36 / 撮影したグリッド数: 1 / 実像数: 2479 / 平均露光時間: 0.2 sec. / 平均電子線量: 46.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 倍率(補正後): 105000 / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 0.01 mm / 最大 デフォーカス(公称値): 0.6 µm / 最小 デフォーカス(公称値): 0.5 µm / 倍率(公称値): 105000 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |