symbiont-mediated arrest of host cell cycle during G2/M transition / permeabilization of host organelle membrane involved in viral entry into host cell / symbiont entry into host cell via permeabilization of inner membrane / viral DNA genome replication / symbiont-mediated perturbation of host cell cycle G1/S transition checkpoint / endonuclease activity / DNA replication / DNA helicase / host cell nucleus / ATP hydrolysis activity ...symbiont-mediated arrest of host cell cycle during G2/M transition / permeabilization of host organelle membrane involved in viral entry into host cell / symbiont entry into host cell via permeabilization of inner membrane / viral DNA genome replication / symbiont-mediated perturbation of host cell cycle G1/S transition checkpoint / endonuclease activity / DNA replication / DNA helicase / host cell nucleus / ATP hydrolysis activity / DNA binding / ATP binding / metal ion binding 類似検索 - 分子機能
Rep protein catalytic-like / : / Rep protein catalytic domain like / Parvovirus (PV) NS1 nuclease (NS1-Nuc) domain profile. / Parvovirus non-structural protein 1, helicase domain / Parvovirus non-structural protein NS1 / Helicase, superfamily 3, DNA virus / Superfamily 3 helicase of DNA viruses domain profile. / P-loop containing nucleoside triphosphate hydrolase 類似検索 - ドメイン・相同性
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM124204
米国
引用
ジャーナル: Nucleic Acids Res / 年: 2020 タイトル: The Cryo-EM structure of AAV2 Rep68 in complex with ssDNA reveals a malleable AAA+ machine that can switch between oligomeric states. 著者: Vishaka Santosh / Faik N Musayev / Rahul Jaiswal / Francisco Zárate-Pérez / Bram Vandewinkel / Caroline Dierckx / Molly Endicott / Kamyar Sharifi / Kelly Dryden / Els Henckaerts / Carlos R Escalante / 要旨: The adeno-associated virus (AAV) non-structural Rep proteins catalyze all the DNA transactions required for virus viability including, DNA replication, transcription regulation, genome packaging, and ...The adeno-associated virus (AAV) non-structural Rep proteins catalyze all the DNA transactions required for virus viability including, DNA replication, transcription regulation, genome packaging, and during the latent phase, site-specific integration. Rep proteins contain two multifunctional domains: an Origin Binding Domain (OBD) and a SF3 helicase domain (HD). Studies have shown that Rep proteins have a dynamic oligomeric behavior where the nature of the DNA substrate molecule modulates its oligomeric state. In the presence of ssDNA, Rep68 forms a large double-octameric ring complex. To understand the mechanisms underlying AAV Rep function, we investigated the cryo-EM and X-ray structures of Rep68-ssDNA complexes. Surprisingly, Rep68 generates hybrid ring structures where the OBD forms octameric rings while the HD forms heptamers. Moreover, the binding to ATPγS promotes a large conformational change in the entire AAA+ domain that leads the HD to form both heptamer and hexamers. The HD oligomerization is driven by an interdomain linker region that acts as a latch to 'catch' the neighboring HD subunit and is flexible enough to permit the formation of different stoichiometric ring structures. Overall, our studies show the structural basis of AAV Rep's structural flexibility required to fulfill its multifunctional role during the AAV life cycle.
ダウンロード / ファイル: emd_22454.map.gz / 形式: CCP4 / 大きさ: 64 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Local refined map of heptameric Rep68 HD complex
ボクセルのサイズ
X=Y=Z: 1.41484 Å
密度
表面レベル
登録者による: 0.413 / ムービー #1: 0.413
最小 - 最大
-2.1726303 - 2.689523
平均 (標準偏差)
-0.0020944097 (±0.089667395)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
0
0
0
サイズ
256
256
256
Spacing
256
256
256
セル
A=B=C: 362.19904 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
1.41483984375
1.41483984375
1.41483984375
M x/y/z
256
256
256
origin x/y/z
0.000
0.000
0.000
length x/y/z
362.199
362.199
362.199
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
93
76
43
NX/NY/NZ
114
126
230
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
256
256
256
D min/max/mean
-2.173
2.690
-0.002
-
添付データ
-
試料の構成要素
-
全体 : Helicase domain heptamer of AAV-2 Rep68 complex bound to AAVS1 ssDNA.
全体
名称: Helicase domain heptamer of AAV-2 Rep68 complex bound to AAVS1 ssDNA.
要素
複合体: Helicase domain heptamer of AAV-2 Rep68 complex bound to AAVS1 ssDNA.
複合体: AAV-2 Rep68
タンパク質・ペプチド: Protein Rep68
複合体: AAVS1 ssDNA
DNA: DNA (5'-D(P*CP*GP*CP*TP*CP*GP*CP*TP*CP*GP*CP*TP*CP*GP*C)-3')
-
超分子 #1: Helicase domain heptamer of AAV-2 Rep68 complex bound to AAVS1 ssDNA.
超分子
名称: Helicase domain heptamer of AAV-2 Rep68 complex bound to AAVS1 ssDNA. タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: all 詳細: Model from local resolution electron density map of helicase domain.
由来(天然)
生物種: Adeno-associated virus - 2 (アデノ随伴ウイルス)
分子量
理論値: 4.481 kDa/nm
-
超分子 #2: AAV-2 Rep68
超分子
名称: AAV-2 Rep68 / タイプ: complex / ID: 2 / 親要素: 1 / 含まれる分子: #1 / 詳細: Local map includes portion of the helicase domain