A motif in the C-terminal domain of phiC31 integrase controls the directionality of recombination.
A motif in the C-terminal domain of phiC31 integrase controls the directionality of recombination.
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DOI：
10.1093/nar/gkn269
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发表时间：
2008-07
影响因子：
14.9
通讯作者：
Smith, Margaret C. M.
中科院分区：
文献类型：
--
作者：
Rowley, Paul A.;Smith, Matthew C. A.;Younger, Ellen;Smith, Margaret C. M.
噬菌体C31编码整合酶，其作用于噬菌体和宿主附着位点attP和attB，以形成侧翼为attL和attR的整合前噬菌体。在没有辅助因子的情况下，CX 31整合酶不能催化attL X attR重组以切除原噬菌体。为了理解方向性的机制，表征了在切除中有活性的突变整合酶。超活性整合酶Int E449 K获得催化attL X attR、attL X attL和attR X attR重组的能力，同时保留重组attP X attB的能力。该突变体的催化缺陷衍生物Int S12 A，E449 K在Int S12 A仅与attP/attB复合的条件下，可与attP/attB、attL/attR、attL/attL和attR/attR形成稳定的复合物。对Int E449 K-attL/attR突触事件的进一步分析揭示了对具有不同attL/attR位点取向的两种预测突触结构之一的偏好。几个氨基酸取代赋予活动过度，包括E449 K，被定位到一个预测的卷曲螺旋基序的C-末端结构域的一面。这项工作表明，在C-末端结构域的C31整合酶的基序控制突触界面的形成，在整合和切除，可能通过直接作用于蛋白质-蛋白质相互作用。
Bacteriophage ϕC31 encodes an integrase, which acts on the phage and host attachment sites, attP and attB, to form an integrated prophage flanked by attL and attR. In the absence of accessory factors, ϕC31 integrase cannot catalyse attL x attR recombination to excise the prophage. To understand the mechanism of directionality, mutant integrases were characterized that were active in excision. A hyperactive integrase, Int E449K, gained the ability to catalyse attL x attR, attL x attL and attR x attR recombination whilst retaining the ability to recombine attP x attB. A catalytically defective derivative of this mutant, Int S12A, E449K, could form stable complexes with attP/attB, attL/attR, attL/attL and attR/attR under conditions where Int S12A only complexed with attP/attB. Further analysis of the Int E449K-attL/attR synaptic events revealed a preference for one of the two predicted synapse structures with different orientations of the attL/attR sites. Several amino acid substitutions conferring hyperactivity, including E449K, were localized to one face of a predicted coiled-coil motif in the C-terminal domain. This work shows that a motif in the C-terminal domain of ϕC31 integrase controls the formation of the synaptic interface in both integration and excision, possibly through a direct role in protein–protein interactions.
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影响因子：
14.9
作者：
Lewis, JA;Hatfull, GF
通讯作者：
Hatfull, GF
DOI：
10.1073/pnas.0711649105
发表时间：
2008-03-04
影响因子：
11.1
作者：
Ghosh, Pallavi;Bibb, Lori A.;Hatfull, Graham F.
通讯作者：
Hatfull, Graham F.
ATTB中影响PHIC31积分酶突触和激活DNA裂解的能力的序列。
影响因子：
14.9
作者：
Gupta, Milind;Till, Rob;Smith, Margaret C M
通讯作者：
Smith, Margaret C M
通过将PHIC31积分酶和CRE重组酶的活性结合的大型和复杂转基因的体内组装。
影响因子：
14.9
作者：
Dafhnis-Calas, F;Xu, ZY;Haines, S;Malla, SK;Smith, MCM;Brown, WRA
通讯作者：
Brown, WRA
影响因子：
3.2
作者：
Crellin, PK;Rood, JI
通讯作者：
Rood, JI