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- PDB-9dch: Single-stranded RNA-mediated PRC2 dimer -

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基本情報

登録情報
データベース: PDB / ID: 9dch
タイトルSingle-stranded RNA-mediated PRC2 dimer
要素
  • (Polycomb protein ...) x 2
  • Isoform 2 of Histone-lysine N-methyltransferase EZH2
  • Protein Jumonji
  • RBAP48
  • TERRAmut RNA
  • Zinc finger protein AEBP2
キーワードGENE REGULATION / PRC2 / RNA / RNP complex / chromatin modifier
機能・相同性
機能・相同性情報


protein localization to pericentric heterochromatin / hepatocyte homeostasis / cellular response to trichostatin A / regulation of gliogenesis / negative regulation of striated muscle cell differentiation / regulation of kidney development / [histone H3]-lysine27 N-trimethyltransferase / response to tetrachloromethane / CAF-1 complex / negative regulation of keratinocyte differentiation ...protein localization to pericentric heterochromatin / hepatocyte homeostasis / cellular response to trichostatin A / regulation of gliogenesis / negative regulation of striated muscle cell differentiation / regulation of kidney development / [histone H3]-lysine27 N-trimethyltransferase / response to tetrachloromethane / CAF-1 complex / negative regulation of keratinocyte differentiation / histone H3K27 trimethyltransferase activity / negative regulation of retinoic acid receptor signaling pathway / cerebellar cortex development / primary miRNA binding / random inactivation of X chromosome / regulatory ncRNA-mediated heterochromatin formation / regulation of adaxial/abaxial pattern formation / skeletal muscle satellite cell maintenance involved in skeletal muscle regeneration / histone H3K27 methyltransferase activity / negative regulation of cardiac muscle cell proliferation / sex chromatin / ubiquitin-modified histone reader activity / positive regulation of cell cycle G1/S phase transition / facultative heterochromatin formation / NuRD complex / NURF complex / regulation of cell fate specification / negative regulation of stem cell population maintenance / genomic imprinting / DNA replication-dependent chromatin assembly / Transcription of E2F targets under negative control by p107 (RBL1) and p130 (RBL2) in complex with HDAC1 / ESC/E(Z) complex / regulation of stem cell differentiation / RSC-type complex / negative regulation of stem cell differentiation / protein-lysine N-methyltransferase activity / Polo-like kinase mediated events / cardiac muscle hypertrophy in response to stress / chromatin silencing complex / Transcription of E2F targets under negative control by DREAM complex / pronucleus / G1 to G0 transition / positive regulation of dendrite development / histone H3 methyltransferase activity / cardiac muscle cell proliferation / DNA methylation-dependent constitutive heterochromatin formation / histone methyltransferase activity / negative regulation of G1/S transition of mitotic cell cycle / lncRNA binding / spinal cord development / ATPase complex / negative regulation of gene expression, epigenetic / Sin3-type complex / synaptic transmission, GABAergic / G1/S-Specific Transcription / positive regulation of stem cell population maintenance / histone methyltransferase complex / Transcriptional Regulation by E2F6 / oligodendrocyte differentiation / RNA Polymerase I Transcription Initiation / histone deacetylase complex / negative regulation of transcription elongation by RNA polymerase II / G0 and Early G1 / negative regulation of cell differentiation / positive regulation of protein serine/threonine kinase activity / : / positive regulation of MAP kinase activity / positive regulation of epithelial to mesenchymal transition / subtelomeric heterochromatin formation / ribonucleoprotein complex binding / pericentric heterochromatin / Transcriptional regulation of brown and beige adipocyte differentiation by EBF2 / Cyclin E associated events during G1/S transition / RNA polymerase II core promoter sequence-specific DNA binding / Cyclin A:Cdk2-associated events at S phase entry / nucleosome binding / keratinocyte differentiation / spleen development / protein localization to chromatin / Regulation of TP53 Activity through Acetylation / positive regulation of GTPase activity / Deposition of new CENPA-containing nucleosomes at the centromere / negative regulation of cytokine production involved in inflammatory response / B cell differentiation / SUMOylation of chromatin organization proteins / negative regulation of cell migration / liver development / thymus development / cellular response to leukemia inhibitory factor / ERCC6 (CSB) and EHMT2 (G9a) positively regulate rRNA expression / central nervous system development / ubiquitin binding / Regulation of PTEN gene transcription / PRC2 methylates histones and DNA / hippocampus development / transcription corepressor binding / Regulation of endogenous retroelements by KRAB-ZFP proteins / Defective pyroptosis / Regulation of endogenous retroelements by Piwi-interacting RNAs (piRNAs) / stem cell differentiation
類似検索 - 分子機能
: / Polycomb protein SUZ12-like, Zn domain / EZH2, SET domain / Polycomb protein, VEFS-Box / VEFS-Box of polycomb protein / Histone-lysine N-methyltransferase EZH1/EZH2 / Polycomb repressive complex 2 subunit EZH1/EZH2, tri-helical domain / Pre-SET CXC domain / : / WD repeat binding protein EZH2 ...: / Polycomb protein SUZ12-like, Zn domain / EZH2, SET domain / Polycomb protein, VEFS-Box / VEFS-Box of polycomb protein / Histone-lysine N-methyltransferase EZH1/EZH2 / Polycomb repressive complex 2 subunit EZH1/EZH2, tri-helical domain / Pre-SET CXC domain / : / WD repeat binding protein EZH2 / Polycomb repressive complex 2 tri-helical domain / CXC domain / Ezh2, MCSS domain / Tesmin/TSO1-like CXC domain / Tesmin/TSO1-like CXC domain / CXC domain / Histone-lysine N-methyltransferase EZH1/2-like / CXC domain profile. / Histone-binding protein RBBP4, N-terminal / Histone-binding protein RBBP4 or subunit C of CAF1 complex / : / : / Zinc finger, C5HC2-type / C5HC2 zinc finger / ARID DNA-binding domain / ARID DNA-binding domain superfamily / ARID/BRIGHT DNA binding domain / ARID domain profile. / BRIGHT, ARID (A/T-rich interaction domain) domain / ARID/BRIGHT DNA binding domain / JmjN domain / jmjN domain / JmjN domain profile. / Small domain found in the jumonji family of transcription factors / SET (Su(var)3-9, Enhancer-of-zeste, Trithorax) domain / SET domain / JmjC domain, hydroxylase / SET domain superfamily / SET domain profile. / SET domain / SANT SWI3, ADA2, N-CoR and TFIIIB'' DNA-binding domains / A domain family that is part of the cupin metalloenzyme superfamily. / JmjC domain / JmjC domain profile. / SANT/Myb domain / zinc finger / Zinc finger C2H2 type domain profile. / Zinc finger C2H2 superfamily / Zinc finger C2H2 type domain signature. / Zinc finger C2H2-type / WD domain, G-beta repeat / G-protein beta WD-40 repeat / WD40 repeat, conserved site / Trp-Asp (WD) repeats signature. / Trp-Asp (WD) repeats profile. / Trp-Asp (WD) repeats circular profile. / WD40 repeats / WD40 repeat / WD40-repeat-containing domain superfamily / WD40/YVTN repeat-like-containing domain superfamily
類似検索 - ドメイン・相同性
RNA / Polycomb protein EED / Histone-binding protein RBBP4 / Polycomb protein SUZ12 / Histone-lysine N-methyltransferase EZH2 / Zinc finger protein AEBP2 / Protein Jumonji
類似検索 - 構成要素
生物種Homo sapiens (ヒト)
手法電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.4 Å
データ登録者Song, J.S. / Kasinath, V.K.
資金援助 米国, 2件
組織認可番号
Howard Hughes Medical Institute (HHMI) 米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R00GM132544 米国
引用ジャーナル: bioRxiv / : 2024
タイトル: Diverse RNA Structures Induce PRC2 Dimerization and Inhibit Histone Methyltransferase Activity.
著者: Jiarui Song / Liqi Yao / Anne R Gooding / Valentin Thron / Vignesh Kasinath / Thomas R Cech
要旨: Methyltransferase PRC2 (Polycomb Repressive Complex 2) introduces histone H3K27 trimethylation, a repressive chromatin mark, to tune the differential expression of genes. PRC2 is precisely regulated ...Methyltransferase PRC2 (Polycomb Repressive Complex 2) introduces histone H3K27 trimethylation, a repressive chromatin mark, to tune the differential expression of genes. PRC2 is precisely regulated by accessory proteins, histone post-translational modifications and, notably, RNA. Research on PRC2-associated RNA has mostly focused on the tight-binding G-quadruplex (G4) RNAs, which inhibit PRC2 enzymatic activity in vitro and in cells. Our recent cryo-EM structure provided a molecular mechanism for G4 RNA inactivating PRC2 via dimerization, but it remained unclear how diverse RNAs associate with and regulate PRC2. Here, we show that a single-stranded G-rich RNA and an atypical G4 structure called pUG-fold unexpectedly also mediate near-identical PRC2 dimerization resulting in inhibition of PRC2 methyltransferase activity. The conformational flexibility of arginine-rich loops within subunits EZH2 and AEBP2 of PRC2 can accommodate diverse RNA secondary structures, resulting in protein-RNA and protein-protein interfaces similar to those observed previously with G4 RNA. Furthermore, we address a recent report that failed to detect PRC2-associated RNAs in living cells by demonstrating the insensitivity of PRC2-RNA interaction to photochemical crosslinking. Our results support the significance of RNA-mediated PRC2 regulation by showing that this interaction is not limited to a single RNA secondary structure, consistent with the broad PRC2 transcriptome containing many G-tract RNAs incapable of folding into G4 structures.
履歴
登録2024年8月26日登録サイト: RCSB / 処理サイト: RCSB
改定 1.02025年3月12日Provider: repository / タイプ: Initial release

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構造の表示

構造ビューア分子:
MolmilJmol/JSmol

ダウンロードとリンク

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集合体

登録構造単位
G: TERRAmut RNA
A: Isoform 2 of Histone-lysine N-methyltransferase EZH2
B: Polycomb protein SUZ12
D: RBAP48
F: Zinc finger protein AEBP2
E: Protein Jumonji
C: Polycomb protein EED
H: Isoform 2 of Histone-lysine N-methyltransferase EZH2
I: Polycomb protein SUZ12
K: RBAP48
M: Zinc finger protein AEBP2
L: Protein Jumonji
J: Polycomb protein EED
ヘテロ分子


分子量 (理論値)分子数
合計 (水以外)674,67627
ポリマ-673,76013
非ポリマー91614
00
1


  • 登録構造と同一
  • 登録者が定義した集合体
  • 根拠: 電子顕微鏡法, not applicable
タイプ名称対称操作
identity operation1_5551

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要素

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タンパク質 , 4種, 8分子 AHDKFMEL

#2: タンパク質 Isoform 2 of Histone-lysine N-methyltransferase EZH2 / ENX-1 / Enhancer of zeste homolog 2 / Lysine N-methyltransferase 6


分子量: 86017.859 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: EZH2, KMT6 / 発現宿主: Trichoplusia ni (イラクサキンウワバ)
参照: UniProt: Q15910, [histone H3]-lysine27 N-trimethyltransferase
#4: タンパク質 RBAP48 / Chromatin assembly factor 1 subunit C / CAF-1 subunit C / Chromatin assembly factor I p48 subunit / ...Chromatin assembly factor 1 subunit C / CAF-1 subunit C / Chromatin assembly factor I p48 subunit / CAF-I 48 kDa subunit / CAF-I p48 / Nucleosome-remodeling factor subunit RBAP48 / Retinoblastoma-binding protein 4 / RBBP-4 / Retinoblastoma-binding protein p48


分子量: 47709.527 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: RBBP4, RBAP48 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q09028
#5: タンパク質 Zinc finger protein AEBP2 / Adipocyte enhancer-binding protein 2 / AE-binding protein 2


分子量: 32881.473 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: AEBP2 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q6ZN18
#6: タンパク質 Protein Jumonji / Jumonji/ARID domain-containing protein 2


分子量: 36021.391 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: JARID2, JMJ / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q92833

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Polycomb protein ... , 2種, 4分子 BICJ

#3: タンパク質 Polycomb protein SUZ12 / Chromatin precipitated E2F target 9 protein / ChET 9 protein / Joined to JAZF1 protein / Suppressor ...Chromatin precipitated E2F target 9 protein / ChET 9 protein / Joined to JAZF1 protein / Suppressor of zeste 12 protein homolog


分子量: 82353.133 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: SUZ12, CHET9, JJAZ1, KIAA0160 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: Q15022
#7: タンパク質 Polycomb protein EED / hEED / Embryonic ectoderm development protein / WD protein associating with integrin cytoplasmic ...hEED / Embryonic ectoderm development protein / WD protein associating with integrin cytoplasmic tails 1 / WAIT-1


分子量: 50267.691 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: EED / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: O75530

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RNA鎖 / 非ポリマー , 2種, 15分子 G

#1: RNA鎖 TERRAmut RNA


分子量: 3257.980 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) Homo sapiens (ヒト)
#8: 化合物
ChemComp-ZN / ZINC ION


分子量: 65.409 Da / 分子数: 14 / 由来タイプ: 合成 / : Zn

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詳細

研究の焦点であるリガンドがあるかN
Has protein modificationY

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実験情報

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実験

実験手法: 電子顕微鏡法
EM実験試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法

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試料調製

構成要素
ID名称タイプEntity IDParent-ID由来
1Single-stranded RNA-mediated dimer of Polycomb Repressive Complex 2COMPLEX#1-#70MULTIPLE SOURCES
2dimer of Polycomb Repressive Complex 2COMPLEX#2-#71RECOMBINANT
3TERRAmut RNACOMPLEX#11SYNTHETIC
由来(天然)
IDEntity assembly-ID生物種Ncbi tax-ID
22Homo sapiens (ヒト)9606
33Homo sapiens (ヒト)9606
由来(組換発現)生物種: Trichoplusia ni (イラクサキンウワバ) / : BTI-Tn-5B1-4
緩衝液pH: 7.9
詳細: RNP complex buffer (25 mM HEPES pH 7.9, 50 mM KCl, 2 mM MgCl2, 10% glycerol, and 1 mM TCEP) EM preparation buffer I (25 mM HEPES pH 7.9, 50 mM KCl, 2.5% glycerol, and 1 mM TCEP) EM ...詳細: RNP complex buffer (25 mM HEPES pH 7.9, 50 mM KCl, 2 mM MgCl2, 10% glycerol, and 1 mM TCEP) EM preparation buffer I (25 mM HEPES pH 7.9, 50 mM KCl, 2.5% glycerol, and 1 mM TCEP) EM preparation buffer II (25 mM HEPES pH 7.9, 50 mM KCl, 2.5% glycerol, 0.01%NP-40, and 1 mM TCEP).
試料包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES
詳細: We used streptavidin-affinity grid preparation method with biotin-labeled RNA at 100 nM concentration. PRC2 was applied in excess at 600 nM.
急速凍結装置: LEICA EM GP / 凍結剤: ETHANE / 湿度: 90 % / 凍結前の試料温度: 281 K / 詳細: 2-3s of single side blotting

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電子顕微鏡撮影

実験機器
モデル: Titan Krios / 画像提供: FEI Company
顕微鏡モデル: FEI TITAN KRIOS
電子銃電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: FLOOD BEAM
電子レンズモード: BRIGHT FIELD / 倍率(補正後): 130000 X / 最大 デフォーカス(公称値): 1900 nm / 最小 デフォーカス(公称値): 500 nm / Cs: 2.7 mm
試料ホルダ凍結剤: NITROGEN
試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER
撮影電子線照射量: 50 e/Å2
フィルム・検出器のモデル: FEI FALCON IV (4k x 4k)
撮影したグリッド数: 2 / 実像数: 14230
詳細: Cryo-EM data was collected using a Titan Krios G3i equipped with a Thermo Fisher Falcon 4 direct-electron detector (DED) camera and a Selectris energy filter set with a 10-eV slit width. Data ...詳細: Cryo-EM data was collected using a Titan Krios G3i equipped with a Thermo Fisher Falcon 4 direct-electron detector (DED) camera and a Selectris energy filter set with a 10-eV slit width. Data acquisition was performed using Thermo Fisher EPU at 130,000x magnification (0.97 A/pixel) with a defocus range of minus 1.9 to minus 0.5 micrometer. Movies were collected in EER format with a total dose of 50 electrons per square angstrom and an exposure time of 5.49 s corresponding to 1323 frames.
電子光学装置エネルギーフィルター名称: TFS Selectris X / エネルギーフィルタースリット幅: 10 eV

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解析

EMソフトウェア
ID名称バージョンカテゴリ
1RELION5粒子像選択
2EPU画像取得
4RELION5CTF補正
7Cootモデルフィッティング
9RELION5初期オイラー角割当
10RELION5最終オイラー角割当
11RELION5分類
12RELION53次元再構成
13PHENIXモデル精密化
画像処理詳細: Gain correction was applied during motion correction using Relion own implementation of the UCSF motioncor2 program
CTF補正詳細: CTFind / タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION
粒子像の選択選択した粒子像数: 3385491 / 詳細: TOPAZ auto picking
3次元再構成解像度: 3.4 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 120658 / 対称性のタイプ: POINT
原子モデル構築詳細: Individual PRC2 protomers were built using cryo-EM maps from the multibody refinement. The coordinates of G-quadruplex RNA-bound PRC2 six-subunit complex (PDB: 8FYH) provided a starting model ...詳細: Individual PRC2 protomers were built using cryo-EM maps from the multibody refinement. The coordinates of G-quadruplex RNA-bound PRC2 six-subunit complex (PDB: 8FYH) provided a starting model from which all the coordinates were adjusted and rebuilt in the new map using COOT. The model of each PRC2 promoter was subjected to global refinement and minimization in real space using PHENIX. These were then subjected to manual inspection and adjustment in COOT, followed by refinement again in PHENIX. TERRAmut RNA model was generated using a 10-nucleotide single-stranded fragment (UGAGUGUGAG) using AlphaFold3 and then docked into our map for the position we designated as the RNA density.
原子モデル構築PDB-ID: 8FYH
Accession code: 8FYH / Source name: PDB / タイプ: experimental model

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