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单链 DNA 结合蛋白荧光融合工具盒的开发。

Development of a single-stranded DNA-binding protein fluorescent fusion toolbox.

机构信息

Department of Biomolecular Chemistry, University of Wisconsin School of Medicine and Public Health, Madison, WI 53706, USA.

Department of Biochemistry, University of Wisconsin - Madison, Madison, WI 53706, USA.

出版信息

Nucleic Acids Res. 2020 Jun 19;48(11):6053-6067. doi: 10.1093/nar/gkaa320.

DOI:10.1093/nar/gkaa320
PMID:32374866
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7293020/
Abstract

Bacterial single-stranded DNA-binding proteins (SSBs) bind single-stranded DNA and help to recruit heterologous proteins to their sites of action. SSBs perform these essential functions through a modular structural architecture: the N-terminal domain comprises a DNA binding/tetramerization element whereas the C-terminus forms an intrinsically disordered linker (IDL) capped by a protein-interacting SSB-Ct motif. Here we examine the activities of SSB-IDL fusion proteins in which fluorescent domains are inserted within the IDL of Escherichia coli SSB. The SSB-IDL fusions maintain DNA and protein binding activities in vitro, although cooperative DNA binding is impaired. In contrast, an SSB variant with a fluorescent protein attached directly to the C-terminus that is similar to fusions used in previous studies displayed dysfunctional protein interaction activity. The SSB-IDL fusions are readily visualized in single-molecule DNA replication reactions. Escherichia coli strains in which wildtype SSB is replaced by SSB-IDL fusions are viable and display normal growth rates and fitness. The SSB-IDL fusions form detectible SSB foci in cells with frequencies mirroring previously examined fluorescent DNA replication fusion proteins. Cells expressing SSB-IDL fusions are sensitized to some DNA damaging agents. The results highlight the utility of SSB-IDL fusions for biochemical and cellular studies of genome maintenance reactions.

摘要

细菌单链 DNA 结合蛋白 (SSB) 结合单链 DNA，并帮助将异源蛋白募集到其作用部位。SSB 通过模块化结构架构执行这些基本功能：N 端结构域包含 DNA 结合/四聚化元件，而 C 端形成内在无序的连接子 (IDL)，由与蛋白相互作用的 SSB-Ct 基序封顶。在这里，我们研究了 SSB-IDL 融合蛋白的活性，其中荧光结构域插入到大肠杆菌 SSB 的 IDL 中。SSB-IDL 融合蛋白在体外保持 DNA 和蛋白结合活性，尽管协同 DNA 结合受到损害。相比之下，与先前研究中使用的融合蛋白相似的直接连接到 C 端的荧光蛋白附着的 SSB 变体显示出功能失调的蛋白相互作用活性。在单分子 DNA 复制反应中，很容易观察到 SSB-IDL 融合蛋白。用 SSB-IDL 融合蛋白替代野生型 SSB 的大肠杆菌菌株是可行的，并且显示出正常的生长速率和适应性。SSB-IDL 融合蛋白在细胞中形成可检测的 SSB 焦点，其频率与之前检查的荧光 DNA 复制融合蛋白相匹配。表达 SSB-IDL 融合蛋白的细胞对一些 DNA 损伤剂敏感。结果突出了 SSB-IDL 融合蛋白在基因组维护反应的生化和细胞研究中的实用性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/2e462504dcff/gkaa320fig9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/8f46bcffcba8/gkaa320fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/aa4bb46f5c91/gkaa320fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/d16aaacc9574/gkaa320fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/49a9ff0c8656/gkaa320fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/0c7220263dce/gkaa320fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/683c7a7af86a/gkaa320fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/c7f126b5ec8e/gkaa320fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/95cfdedfea0f/gkaa320fig8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/2e462504dcff/gkaa320fig9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/8f46bcffcba8/gkaa320fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/aa4bb46f5c91/gkaa320fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/d16aaacc9574/gkaa320fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/49a9ff0c8656/gkaa320fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/0c7220263dce/gkaa320fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/683c7a7af86a/gkaa320fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/c7f126b5ec8e/gkaa320fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/95cfdedfea0f/gkaa320fig8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ecb9/7293020/2e462504dcff/gkaa320fig9.jpg

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Biophys J. 2019 Dec 3;117(11):2120-2140. doi: 10.1016/j.bpj.2019.09.047. Epub 2019 Oct 28.
2
Super-resolution imaging reveals changes in Escherichia coli SSB localization in response to DNA damage.超分辨率成像揭示了 DNA 损伤响应中大肠杆菌 SSB 定位的变化。
Genes Cells. 2019 Dec;24(12):814-826. doi: 10.1111/gtc.12729. Epub 2019 Nov 14.
3
Are the intrinsically disordered linkers involved in SSB binding to accessory proteins?
细菌表达克隆筛选发现单链 DNA 结合蛋白是有效的干燥保护剂。
Cell Rep. 2024 Nov 26;43(11):114956. doi: 10.1016/j.celrep.2024.114956. Epub 2024 Nov 11.
4
Mapping fast DNA polymerase exchange during replication.绘制复制过程中快速 DNA 聚合酶交换图谱。
Nat Commun. 2024 Jun 22;15(1):5328. doi: 10.1038/s41467-024-49612-3.
5
Molecular insights into the prototypical single-stranded DNA-binding protein from .从. 中获得的典型单链 DNA 结合蛋白的分子见解。
Crit Rev Biochem Mol Biol. 2024 Feb-Apr;59(1-2):99-127. doi: 10.1080/10409238.2024.2330372. Epub 2024 May 21.
6
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7
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Cell Death Discov. 2023 Aug 28;9(1):319. doi: 10.1038/s41420-023-01619-3.
8
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