IRON/SULFUR CLUSTER / DNA / DNA (> 10) / DNA (> 100) / : / : / DNA-directed RNA polymerase II subunit RPB1 / General transcription and DNA repair factor IIH helicase subunit XPD/RAD3 / DNA-directed RNA polymerase II subunit RPB2 / TATA-box-binding protein ...IRON/SULFUR CLUSTER / DNA / DNA (> 10) / DNA (> 100) / : / : / DNA-directed RNA polymerase II subunit RPB1 / General transcription and DNA repair factor IIH helicase subunit XPD/RAD3 / DNA-directed RNA polymerase II subunit RPB2 / TATA-box-binding protein / DNA-directed RNA polymerase II subunit RPB3 / Mediator of RNA polymerase II transcription subunit 14 / DNA-directed RNA polymerase II subunit RPB4 / DNA-directed RNA polymerases I, II, and III subunit RPABC1 / DNA-directed RNA polymerases I, II, and III subunit RPABC2 / DNA-directed RNA polymerases I, II, and III subunit RPABC3 / DNA-directed RNA polymerases I, II, and III subunit RPABC5 / Mediator of RNA polymerase II transcription subunit 19 / DNA-directed RNA polymerase II subunit RPB9 / Transcription initiation factor IIB / Mediator of RNA polymerase II transcription subunit 17 / Mediator of RNA polymerase II transcription subunit 22 / Mediator of RNA polymerase II transcription subunit 18 / Transcription initiation factor IIA large subunit / Transcription initiation factor IIA subunit 2 / General transcription and DNA repair factor IIH subunit TFB1 / Mediator of RNA polymerase II transcription subunit 9 / DNA-directed RNA polymerase II subunit RPB7 / Mediator of RNA polymerase II transcription subunit 20 / Transcription initiation factor IIE subunit alpha / Transcription initiation factor IIE subunit beta / Mediator of RNA polymerase II transcription subunit 8 / Mediator of RNA polymerase II transcription subunit 31 / Mediator of RNA polymerase II transcription subunit 6 / DNA-directed RNA polymerase II subunit RPB11 / DNA-directed RNA polymerases I, II, and III subunit RPABC4 / Transcription initiation factor IIF subunit alpha / Transcription initiation factor IIF subunit beta / Mediator of RNA polymerase II transcription subunit 21 / General transcription and DNA repair factor IIH helicase/translocase subunit XPB/SSL2 / General transcription and DNA repair factor IIH subunit TFB2 / RNA polymerase II transcription factor B subunit 3 / General transcription and DNA repair factor IIH subunit SSL1 / Mediator of RNA polymerase II transcription subunit 10 / Mediator of RNA polymerase II transcription subunit 7 / General transcription and DNA repair factor IIH subunit TFB4 / Mediator of RNA polymerase II transcription subunit 4 / General transcription and DNA repair factor IIH subunit TFB5 / Mediator of RNA polymerase II transcription subunit 11 類似検索 - 構成要素
ジャーナル: Nature / 年: 2017 タイトル: Structures of transcription pre-initiation complex with TFIIH and Mediator. 著者: S Schilbach / M Hantsche / D Tegunov / C Dienemann / C Wigge / H Urlaub / P Cramer / 要旨: For the initiation of transcription, RNA polymerase II (Pol II) assembles with general transcription factors on promoter DNA to form the pre-initiation complex (PIC). Here we report cryo-electron ...For the initiation of transcription, RNA polymerase II (Pol II) assembles with general transcription factors on promoter DNA to form the pre-initiation complex (PIC). Here we report cryo-electron microscopy structures of the Saccharomyces cerevisiae PIC and PIC-core Mediator complex at nominal resolutions of 4.7 Å and 5.8 Å, respectively. The structures reveal transcription factor IIH (TFIIH), and suggest how the core and kinase TFIIH modules function in the opening of promoter DNA and the phosphorylation of Pol II, respectively. The TFIIH core subunit Ssl2 (a homologue of human XPB) is positioned on downstream DNA by the 'E-bridge' helix in TFIIE, consistent with TFIIE-stimulated DNA opening. The TFIIH kinase module subunit Tfb3 (MAT1 in human) anchors the kinase Kin28 (CDK7), which is mobile in the PIC but preferentially located between the Mediator hook and shoulder in the PIC-core Mediator complex. Open spaces between the Mediator head and middle modules may allow access of the kinase to its substrate, the C-terminal domain of Pol II.
A: DNA-directed RNA polymerase II subunit RPB1 B: DNA-directed RNA polymerase II subunit RPB2 C: DNA-directed RNA polymerase II subunit RPB3 D: DNA-directed RNA polymerase II subunit RPB4 E: DNA-directed RNA polymerases I, II, and III subunit RPABC1 F: DNA-directed RNA polymerases I, II, and III subunit RPABC2 G: DNA-directed RNA polymerase II subunit RPB7 H: DNA-directed RNA polymerases I, II, and III subunit RPABC3 I: DNA-directed RNA polymerase II subunit RPB9 J: DNA-directed RNA polymerases I, II, and III subunit RPABC5 K: DNA-directed RNA polymerase II subunit RPB11 L: DNA-directed RNA polymerases I, II, and III subunit RPABC4 M: Transcription initiation factor IIB N: NONTEMPLATE DNA O: TATA-box-binding protein Q: Transcription initiation factor IIF subunit alpha R: Transcription initiation factor IIF subunit beta T: TEMPLATE DNA U: Transcription initiation factor IIA large subunit V: Transcription initiation factor IIA subunit 2 W: Transcription factor TFIIE subunit X: Transcription initiation factor IIE subunit beta 0: General transcription and DNA repair factor IIH helicase subunit XPD 1: General transcription and DNA repair factor IIH subunit TFB1 2: General transcription and DNA repair factor IIH subunit TFB2 3: RNA polymerase II transcription factor B subunit 3 4: General transcription and DNA repair factor IIH subunit TFB4 5: General transcription and DNA repair factor IIH subunit TFB5 6: General transcription and DNA repair factor IIH subunit SSL1 7: General transcription and DNA repair factor IIH helicase subunit XPB Z: Unknown protein a: Mediator of RNA polymerase II transcription subunit 6 b: Mediator of RNA polymerase II transcription subunit 8 c: Mediator of RNA polymerase II transcription subunit 11 d: Mediator of RNA polymerase II transcription subunit 17 e: Mediator of RNA polymerase II transcription subunit 18 f: Mediator of RNA polymerase II transcription subunit 20 g: Mediator of RNA polymerase II transcription subunit 22 h: Mediator of RNA polymerase II transcription subunit 4 i: Mediator of RNA polymerase II transcription subunit 7 j: Mediator of RNA polymerase II transcription subunit 9 k: Mediator of RNA polymerase II transcription subunit 10 l: Mediator of RNA polymerase II transcription subunit 14 m: Mediator of RNA polymerase II transcription subunit 19 n: Mediator of RNA polymerase II transcription subunit 21 o: Mediator of RNA polymerase II transcription subunit 31,Mediator of RNA polymerase II transcription subunit 31 ヘテロ分子
DNA-directed RNA polymerase II subunit ... , 7種, 7分子 ABCDGIK
#1: タンパク質
DNA-directedRNApolymeraseIIsubunitRPB1 / RNA polymerase II subunit B1 / DNA-directed RNA polymerase III largest subunit / RNA polymerase II ...RNA polymerase II subunit B1 / DNA-directed RNA polymerase III largest subunit / RNA polymerase II subunit B220
DNA-directed RNA polymerases I, II, and III subunit ... , 5種, 5分子 EFHJL
#5: タンパク質
DNA-directedRNApolymerasesI, II, andIIIsubunitRPABC1 / RNA polymerases I / II / and III subunit ABC1 / ABC27 / DNA-directed RNA polymerases I / and III 27 ...RNA polymerases I / II / and III subunit ABC1 / ABC27 / DNA-directed RNA polymerases I / and III 27 kDa polypeptide
DNA-directedRNApolymerasesI, II, andIIIsubunitRPABC2 / RNA polymerases I / II / and III subunit ABC2 / ABC23 / DNA-directed RNA polymerases I / and III 23 ...RNA polymerases I / II / and III subunit ABC2 / ABC23 / DNA-directed RNA polymerases I / and III 23 kDa polypeptide
DNA-directedRNApolymerasesI, II, andIIIsubunitRPABC3 / RNA polymerases I / II / and III subunit ABC3 / ABC14.4 / ABC14.5 / DNA-directed RNA polymerases I ...RNA polymerases I / II / and III subunit ABC3 / ABC14.4 / ABC14.5 / DNA-directed RNA polymerases I / and III 14.5 kDa polypeptide
DNA-directedRNApolymerasesI, II, andIIIsubunitRPABC5 / RNA polymerases I / II / and III subunit ABC5 / ABC10-beta / ABC8 / DNA-directed RNA polymerases I ...RNA polymerases I / II / and III subunit ABC5 / ABC10-beta / ABC8 / DNA-directed RNA polymerases I / and III 8.3 kDa polypeptide
RNApolymeraseIItranscriptionfactorBsubunit3 / RNA polymerase II transcription factor B 38 kDa subunit / RNA polymerase II transcription factor B p38 subunit
Residues indicated as UNK are of uncertain amino acid register and may deviate from the given ...Residues indicated as UNK are of uncertain amino acid register and may deviate from the given register by few amino acids. For details about modelling of the subunits refer to the associated publication. Residues indicated as UNK with the chain identifier Z are part of an assembly between the clutch domains of Tfb2 and Ssl2 but could not be assigned to any of the two proteins with confidence since domain swapping/beta-strand exchange may occur. To account for this uncertainty, a theoretical chain Z was introduced for this residue range but not assigned to any of the PIC components. Submission based on experimental data from EMDB EMD-3850.
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実験情報
-
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 ID
Parent-ID
由来
1
Pre-Initiation Complex with core Mediator (PIC-cMed)
COMPLEX
#1-#46
0
MULTIPLESOURCES
2
DNA-directed RNA polymerase
COMPLEX
#1-#12, #31
1
NATURAL
3
RNApolymeraseIItranscriptionfactorB
COMPLEX
#13, #15-#17, #19-#22, #32-#46
1
RECOMBINANT
4
MediatorofRNApolymeraseIItranscription
COMPLEX
#23-#30
1
RECOMBINANT
5
DNA
COMPLEX
#14, #18
1
RECOMBINANT
分子量
値: 1.95 MDa / 実験値: NO
由来(天然)
ID
Entity assembly-ID
生物種
Ncbi tax-ID
1
2
Saccharomyces cerevisiae (パン酵母)
4932
2
3
Saccharomyces cerevisiae (パン酵母)
4932
3
4
Saccharomyces cerevisiae (パン酵母)
4932
4
5
synthetic construct (人工物)
32630
由来(組換発現)
ID
Entity assembly-ID
生物種
Ncbi tax-ID
1
3
Escherichia coli BL21(DE3) (大腸菌)
469008
2
4
Trichoplusia ni (イラクサキンウワバ)
7111
3
5
synthetic construct (人工物)
32630
緩衝液
pH: 7.6
試料
濃度: 0.7 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES
試料支持
詳細: QUANTIFOIL HOLEY GOLD GRIDS WERE GLOW-DISCHARGED BEFORE DEPOSITION OF SAMPLE. グリッドの材料: GOLD / グリッドのタイプ: Quantifoil R1.2/1.3
急速凍結
装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE 詳細: Grids were blotted for 8 s and plunge-frozen in liquid ethane.