Examination of an inverted repeat within the F factor origin of transfer: context dependence of F TraI relaxase DNA specificity.
Examination of an inverted repeat within the F factor origin of transfer: context dependence of F TraI relaxase DNA specificity.
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DOI：
10.1093/nar/gkj444
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发表时间：
2006
影响因子：
14.9
通讯作者：
Schildbach, JF
中科院分区：
文献类型：
--
作者：
Williams, SL;Schildbach, JF
在质粒接合转移之前，一条质粒链被称为松弛酶或切口酶的酶以位点和链特异性方式切割。在F和相关质粒中，反向重复位于质粒链切割位点附近，其他人提出该序列在单链形式时形成发夹的能力对于转移很重要。将取代引入克隆的F oriT 区域并评估它们对质粒转移的影响。对于那些显着减少转移的取代，结果通常与寡核苷酸与F TraI松弛酶结构域的体外结合的影响相关，而不是与发夹形成的预测影响相关。先前显示的一种取代可显着减少质粒转移和与 17 碱基寡核苷酸的体外结合，但对与包含发夹区域的 30 碱基寡核苷酸的结合几乎没有明显影响。随后实验的结果强烈表明，松弛酶结构域可以以具有不同序列特异性的两种不同方式结合发夹寡核苷酸，并且该蛋白质在相同或重叠位点结合寡核苷酸。
Prior to conjugative transfer of plasmids, one plasmid strand is cleaved in a site- and strand-specific manner by an enzyme called a relaxase or nickase. In F and related plasmids, an inverted repeat is located near the plasmid strand cleavage site, and others have proposed that the ability of this sequence to form a hairpin when in single-stranded form is important for transfer. Substitutions were introduced into a cloned F oriT region and their effects on plasmid transfer were assessed. For those substitutions that substantially reduced transfer, the results generally correlated with effects on in vitro binding of oligonucleotides to the F TraI relaxase domain rather than with predicted effects on hairpin formation. One substitution shown previously to dramatically reduce both plasmid transfer and in vitro binding to a 17-base oligonucleotide had little apparent effect on binding to a 30-base oligonucleotide that contained the hairpin region. Results from subsequent experiments strongly suggest that the relaxase domain can bind to hairpin oligonucleotides in two distinct manners with different sequence specificities, and that the protein binds the oligonucleotides at the same or overlapping sites.
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影响因子：
16
作者：
Hickman, AB;Ronning, DR;Dyda, F
通讯作者：
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影响因子：
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作者：
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DOI：
10.1073/pnas.90.7.2925
发表时间：
1993-04-01
影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
THOMPSON, R;TAYLOR, L;WILLETTS, N
通讯作者：
WILLETTS, N
DOI：
10.1073/pnas.0506081102
发表时间：
2005-11-08
影响因子：
11.1
作者：
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通讯作者：
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