Conserved residues of the C-terminal p16 domain of primase are involved in modulating the activity of the bacterial primosome.
Conserved residues of the C-terminal p16 domain of primase are involved in modulating the activity of the bacterial primosome.
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DOI：
10.1111/j.1365-2958.2008.06155.x
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发表时间：
2008-04
影响因子：
3.6
通讯作者：
Soultanas P
中科院分区：
文献类型：
--
作者：
Chintakayala K;Larson MA;Griep MA;Hinrichs SH;Soultanas P
细菌引发体包含复制性同源六聚环解旋酶DnaB和引发酶DnaG。它是复制体的一个组成部分，因为它解开亲本DNA双链体以允许复制叉的进展，在复制起点合成起始引物oriC，以及在滞后链复制期间冈崎片段合成所需的引物。两种组分蛋白之间的相互作用由引发酶的不同C-末端结构域（p16）介导。这两种蛋白质相互调节对方的活动和保守残基的推定网络已被提出来介导这些影响。我们已经从这个网络中锁定了10个残基。为了研究这些残基对引发酶、ATP酶和解旋酶活性的功能贡献，我们使用了定点诱变和体外功能测定。这些残基中的五个（E464、H494、R495、Y 548和R555）表现出一些功能意义，而其余五个（E483、R484、E506、D512和E530）表现出无影响。E464参与引发酶活性的功能调节，而H494、R495和R555参与ATP酶和/或解旋酶活性的变构功能调节。Y 548直接参与与DnaB的结构相互作用。
The bacterial primosome comprises the replicative homo-hexameric ring helicase DnaB and the primase DnaG. It is an integral component of the replisome as it unwinds the parental DNA duplex to allow progression of the replication fork, synthesizes the initiation primers at the replication origin, oriC, and the primers required for Okazaki fragment synthesis during lagging strand replication. The interaction between the two component proteins is mediated by a distinct C-terminal domain (p16) of the primase. Both proteins mutually regulate each other’s activities and a putative network of conserved residues has been proposed to mediate these effects. We have targeted 10 residues from this network. To investigate the functional contributions of these residues to the primase, ATPase and helicase activities of the primosome, we have used site-directed mutagenesis and in vitro functional assays. Five of these residues (E464, H494, R495, Y548 and R555) exhibited some functional significance while the remaining five (E483, R484, E506, D512 and E530) exhibited no effects. E464 participates in functional modulation of the primase activity, whereas H494, R495 and R555 participate in allosteric functional modulation of the ATPase and/or helicase activities. Y548 contributes directly to the structural interaction with DnaB.
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影响因子：
14.9
作者：
Soultanas, P;Wigley, DB
通讯作者：
Wigley, DB
影响因子：
5.7
作者：
Pan, H;Wigley, DB
通讯作者：
Wigley, DB
影响因子：
64.8
作者：
Lee, JB;Hite, RK;van Oijen, AM
通讯作者：
van Oijen, AM
DOI：
10.1016/j.str.2005.01.022
发表时间：
2005-04
期刊：
Structure (London, England : 1993)
影响因子：
--
作者：
Syson K;Thirlway J;Hounslow AM;Soultanas P;Waltho JP
通讯作者：
Waltho JP
影响因子：
4.8
作者：
Mitkova, AV;Khopde, SM;Biswas, SB
通讯作者：
Biswas, SB