- EMDB-30366: Human DMC1 Q244M mutant of the post-synaptic complexes -
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基本情報
登録情報
データベース: EMDB / ID: EMD-30366
タイトル
Human DMC1 Q244M mutant of the post-synaptic complexes
マップデータ
試料
複合体: DMC1Q244M-dsDNA filament
複合体: DMC1 Q244M mutant
タンパク質・ペプチド: Meiotic recombination protein DMC1/LIM15 homolog
複合体: homologous double strand DNA
リガンド: CALCIUM ION
リガンド: PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER
キーワード
meiotic homologous recombination / DNA repair / ATPase / RECOMBINATION
機能・相同性
機能・相同性情報
female gamete generation / chromosome organization involved in meiotic cell cycle / double-strand break repair involved in meiotic recombination / homologous chromosome pairing at meiosis / lateral element / DNA recombinase assembly / mitotic recombination / DNA strand invasion / DNA strand exchange activity / reciprocal meiotic recombination ...female gamete generation / chromosome organization involved in meiotic cell cycle / double-strand break repair involved in meiotic recombination / homologous chromosome pairing at meiosis / lateral element / DNA recombinase assembly / mitotic recombination / DNA strand invasion / DNA strand exchange activity / reciprocal meiotic recombination / oocyte maturation / ATP-dependent DNA damage sensor activity / male meiosis I / spermatid development / ATP-dependent activity, acting on DNA / ovarian follicle development / condensed nuclear chromosome / meiotic cell cycle / Meiotic recombination / chromosome / site of double-strand break / single-stranded DNA binding / spermatogenesis / double-stranded DNA binding / chromosome, telomeric region / ATP hydrolysis activity / DNA binding / ATP binding / identical protein binding / nucleus 類似検索 - 分子機能
Meiotic recombination protein Dmc1 / DNA recombination and repair protein, RecA-like / DNA recombination and repair protein Rad51-like, C-terminal / Rad51 / DNA recombination and repair protein RecA, monomer-monomer interface / RecA family profile 2. / DNA recombination and repair protein RecA-like, ATP-binding domain / RecA family profile 1. / DNA repair Rad51/transcription factor NusA, alpha-helical / Helix-hairpin-helix domain ...Meiotic recombination protein Dmc1 / DNA recombination and repair protein, RecA-like / DNA recombination and repair protein Rad51-like, C-terminal / Rad51 / DNA recombination and repair protein RecA, monomer-monomer interface / RecA family profile 2. / DNA recombination and repair protein RecA-like, ATP-binding domain / RecA family profile 1. / DNA repair Rad51/transcription factor NusA, alpha-helical / Helix-hairpin-helix domain / ATPases associated with a variety of cellular activities / AAA+ ATPase domain / P-loop containing nucleoside triphosphate hydrolase 類似検索 - ドメイン・相同性
Meiotic recombination protein DMC1/LIM15 homolog 類似検索 - 構成要素
ジャーナル: Nat Commun / 年: 2021 タイトル: Identification of fidelity-governing factors in human recombinases DMC1 and RAD51 from cryo-EM structures. 著者: Shih-Chi Luo / Hsin-Yi Yeh / Wei-Hsuan Lan / Yi-Min Wu / Cheng-Han Yang / Hao-Yen Chang / Guan-Chin Su / Chia-Yi Lee / Wen-Jin Wu / Hung-Wen Li / Meng-Chiao Ho / Peter Chi / Ming-Daw Tsai / 要旨: Both high-fidelity and mismatch-tolerant recombination, catalyzed by RAD51 and DMC1 recombinases, respectively, are indispensable for genomic integrity. Here, we use cryo-EM, MD simulation and ...Both high-fidelity and mismatch-tolerant recombination, catalyzed by RAD51 and DMC1 recombinases, respectively, are indispensable for genomic integrity. Here, we use cryo-EM, MD simulation and functional analysis to elucidate the structural basis for the mismatch tolerance of DMC1. Structural analysis of DMC1 presynaptic and postsynaptic complexes suggested that the lineage-specific Loop 1 Gln244 (Met243 in RAD51) may help stabilize DNA backbone, whereas Loop 2 Pro274 and Gly275 (Val273/Asp274 in RAD51) may provide an open "triplet gate" for mismatch tolerance. In support, DMC1-Q244M displayed marked increase in DNA dynamics, leading to unobservable DNA map. MD simulation showed highly dispersive mismatched DNA ensemble in RAD51 but well-converged DNA in DMC1 and RAD51-V273P/D274G. Replacing Loop 1 or Loop 2 residues in DMC1 with RAD51 counterparts enhanced DMC1 fidelity, while reciprocal mutations in RAD51 attenuated its fidelity. Our results show that three Loop 1/Loop 2 residues jointly enact contrasting fidelities of DNA recombinases.
名称: homologous double strand DNA / タイプ: complex / ID: 3 / 親要素: 1 / 含まれる分子: #1 詳細: density map of DNA is missing duo to the flexible/dynamic property of dsDNA in the DMC1 Q244M filament
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分子 #1: Meiotic recombination protein DMC1/LIM15 homolog
凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 295 K / 装置: FEI VITROBOT MARK IV 詳細: The grids were blotted for 1 sec at 22 degree C with 100% relative humidity and plunge-frozen in liquid ethane cooled by liquid nitrogen using a Vitrobot Mark IV (Thermo Fisher)..
詳細
protein sample were applied onto a pre-glow-discharged graphene-oxide coated Quantifoil holey carbon grid (1.2/1.3, 200 mesh) using published protocol
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
アライメント法
Coma free - Residual tilt: 10.0 mrad
特殊光学系
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: 30 eV
選択した数: 643100 / ソフトウェア - 名称: RELION (ver. 3.0) 詳細: Filaments were manually picked, and segments were extracted using a box size of 384 pixel and an inter-box distance of ~10% of the box length.
初期モデル
モデルのタイプ: INSILICO MODEL In silico モデル: A simple cylinder was used as initial model to prevent model bias.