The DNA-binding domain of human PARP-1 interacts with DNA single-strand breaks as a monomer through its second zinc finger.
The DNA-binding domain of human PARP-1 interacts with DNA single-strand breaks as a monomer through its second zinc finger.
复制标题
DOI：
10.1016/j.jmb.2011.01.034
复制
发表时间：
2011-03-18
影响因子：
5.6
通讯作者：
Neuhaus D
中科院分区：
文献类型：
--
作者：
Eustermann S;Videler H;Yang JC;Cole PT;Gruszka D;Veprintsev D;Neuhaus D
多聚腺苷二磷酸核糖聚合酶-1（Poly（ADP-ribose）polymerase-1，PARP-1）是一种存在于所有高等真核细胞核中的染色质相关酶，在维持基因组完整性、染色质重塑和转录调控等方面发挥重要作用。它通过含有两个锌指F1和F2的N-末端区域与DNA单链和双链断裂结合，随后其C-末端催化结构域通过未知机制被激活，导致聚腺苷核糖（PAR）形成并添加到受体蛋白（包括PARP-1本身）。在这里，我们报告的F1和F2指的人PARP-1的生物物理和结构表征，无论是作为独立的片段，并在上下文中的24 kDa的DNA结合域（F1 + F2）。我们表明，手指是结构上独立的DNA的情况下，共享一个高度相似的结构折叠和动力学。F1 + F2片段将DNA单链断裂识别为单体并以单一方向。使用NMR光谱和其他生物物理技术的组合，我们表明，识别主要是通过F2，它结合的DNA在一个基本上相同的方式是否存在于隔离或在两个手指片段。F2与有缺口或有缺口的DNA配体的相互作用比F1强得多，我们提出了一个突变研究，表明这种差异的起源。我们的数据表明，不同的DNA损伤识别的DNA结合结构域的PARP-1在一个高度相似的构象，有助于合理化如何全长蛋白质参与多个步骤的DNA单链断裂和碱基切除修复。PARP-1锌指1和2在自由状态下是结构独立的。CAPARP-1指1 + 2形成1：1单体复合物，具有DNA单链断裂。PARP-1的1 + 2指类似地识别不同类型的DNA单链断裂。PARP-1对DNA损伤的识别主要通过第2指进行。
Poly(ADP-ribose)polymerase-1 (PARP-1) is a highly abundant chromatin-associated enzyme present in all higher eukaryotic cell nuclei, where it plays key roles in the maintenance of genomic integrity, chromatin remodeling and transcriptional control. It binds to DNA single- and double-strand breaks through an N-terminal region containing two zinc fingers, F1 and F2, following which its C-terminal catalytic domain becomes activated via an unknown mechanism, causing formation and addition of polyadenosine-ribose (PAR) to acceptor proteins including PARP-1 itself. Here, we report a biophysical and structural characterization of the F1 and F2 fingers of human PARP-1, both as independent fragments and in the context of the 24-kDa DNA-binding domain (F1 + F2). We show that the fingers are structurally independent in the absence of DNA and share a highly similar structural fold and dynamics. The F1 + F2 fragment recognizes DNA single-strand breaks as a monomer and in a single orientation. Using a combination of NMR spectroscopy and other biophysical techniques, we show that recognition is primarily achieved by F2, which binds the DNA in an essentially identical manner whether present in isolation or in the two-finger fragment. F2 interacts much more strongly with nicked or gapped DNA ligands than does F1, and we present a mutational study that suggests origins of this difference. Our data suggest that different DNA lesions are recognized by the DNA-binding domain of PARP-1 in a highly similar conformation, helping to rationalize how the full-length protein participates in multiple steps of DNA single-strand breakage and base excision repair. ► PARP-1 zinc fingers 1 and 2 are structurally independent in the free state. ► PARP-1 fingers 1 + 2 form a 1:1 monomeric complex with DNA single-strand breaks. ► PARP-1 fingers 1 + 2 recognize different types of DNA single-strand break similarly. ► DNA damage recognition by PARP-1 occurs predominantly through finger 2.
登录
查看更多内容
影响因子：
14.9
作者：
Altmeyer M;Messner S;Hassa PO;Fey M;Hottiger MO
通讯作者：
Hottiger MO
DOI：
10.1107/s0907444994010723
发表时间：
1995-03-01
期刊：
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
影响因子：
--
作者：
DIAMOND, R
通讯作者：
DIAMOND, R
影响因子：
21.3
作者：
Kanai, Masayuki;Hanashiro, Kazuhiko;Fukasawa, Kenji
通讯作者：
Fukasawa, Kenji
影响因子：
9.2
作者：
Jeggo, PA
通讯作者：
Jeggo, PA
影响因子：
16.8
作者：
通讯作者：
--