Nucleoprotein architectures regulating the directionality of viral integration and excision

Nicole E. Seah; David Warren; Wenjun TongGurunathan LaxmikanthanGregory D. Van DuyneArthur Landya

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Nucleoprotein architectures regulating the directionality of viral integration and excision

机译：Nucleoprotein architectures regulating the directionality of viral integration and excision

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The virally encoded site-specific recombinase Int collaborates with its accessory DNA bending proteins IHF, Xis, and Fis to assemble two distinct, very large, nucleoprotein complexes that carry out either integrative or excisive recombination along regulated and essentially unidirectional pathways. The core of each complex consists of a tetramer of Integrase protein (Int), which is a heterobivalent DNA binding protein that binds and bridges a core-type DNA site (where strand cleavage and ligation are executed), and a distal arm-type site, that is brought within range by one or more DNA bending proteins. The recent determination of the patterns of these Int bridges has made it possible to think realistically about the global architecture of the recombinogenic complexes. Here, we combined the previously determined Int bridging patterns with in-gel FRET experiments and in silico modeling to characterize and differentiate the two 400-kDa multiprotein Holiday junction recombination intermediates formed during λ integration and excision. The results lead to architectural models that explain how integration and excision are regulated in λ site-specific recombination. Our confidence in the basic features of these architectures is based on the redundancy and self-consistency of the underlying data from two very different experimental approaches to establish bridging interactions, a set of strategic intracomplex distances from FRET experiments, and the model’s ability to explain key aspects of the integrative and excisive recombination pathways, such as topological changes, the mechanism of capturing attB, and the features of asymmetry and flexibility within the complexes.

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《Proceedings of the National Academy of Sciences of the United States of America》 |2014年第34期|12372-12377|共6页
作者
Nicole E. Seah; David Warren; Wenjun TongGurunathan LaxmikanthanGregory D. Van DuyneArthur Landya;
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作者单位

Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104;

Department of Molecular Biology, Cell Biology, and Biochemistry, Division of Biology and Medicine, Brown University, Providence, RI 02912;

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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
原文格式 PDF
正文语种英语
中图分类自然科学总论;
关键词
regulation of directionality; topology; recombinogenic architectures; molecular machines;

Nucleoprotein architectures regulating the directionality of viral integration and excision

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