{"title":"肠致病性大肠杆菌中Ler的DNA结合域与DNA复合物的1H, 13C和15N共振分配","authors":"Fan Liu, Bo Duan, Bin Xia","doi":"10.1007/s00723-023-01575-2","DOIUrl":null,"url":null,"abstract":"<div><p>Ler is a master regulator for the gene regulation of the locus of enterocyte effacement (LEE) pathogenicity island (PAI), which could activate the transcription of <i>LEE2</i> to <i>LEE5</i> by counteracting the repression of H-NS. Ler contains an N-terminal oligomerization domain, a linker region, and a C-terminal DNA binding domain. However, the DNA binding mechanism of Ler remains unclear. Here, we report the <sup>1</sup>H, <sup>13</sup>C, and <sup>15</sup>N NMR assignments of Ler<sup>CTD</sup>/3A3T-DNA complex. We have achieved 97.8% completeness for the backbone and side-chain resonance assignments of Ler<sup>CTD</sup> in the complex, excluding <sup>13</sup>C′ resonances. In addition, about 91.5% of the <sup>1</sup>H resonances of 3A3T-DNA have been assigned. Our near complete assignments of this complex will significantly advance the parsing of the intermolecular NOE signals between Ler <sup>CTD</sup> and 3A3T-DNA. Therefore, our NMR assignments formed a good basis for further DNA binding mechanism studies of Ler.</p></div>","PeriodicalId":469,"journal":{"name":"Applied Magnetic Resonance","volume":"54 10","pages":"979 - 985"},"PeriodicalIF":1.1000,"publicationDate":"2023-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00723-023-01575-2.pdf","citationCount":"0","resultStr":"{\"title\":\"1H, 13C, and 15N Resonance Assignments of the DNA Binding Domain of Ler from Enteropathogenic E. coli in Complex with DNA\",\"authors\":\"Fan Liu, Bo Duan, Bin Xia\",\"doi\":\"10.1007/s00723-023-01575-2\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Ler is a master regulator for the gene regulation of the locus of enterocyte effacement (LEE) pathogenicity island (PAI), which could activate the transcription of <i>LEE2</i> to <i>LEE5</i> by counteracting the repression of H-NS. Ler contains an N-terminal oligomerization domain, a linker region, and a C-terminal DNA binding domain. However, the DNA binding mechanism of Ler remains unclear. Here, we report the <sup>1</sup>H, <sup>13</sup>C, and <sup>15</sup>N NMR assignments of Ler<sup>CTD</sup>/3A3T-DNA complex. We have achieved 97.8% completeness for the backbone and side-chain resonance assignments of Ler<sup>CTD</sup> in the complex, excluding <sup>13</sup>C′ resonances. In addition, about 91.5% of the <sup>1</sup>H resonances of 3A3T-DNA have been assigned. Our near complete assignments of this complex will significantly advance the parsing of the intermolecular NOE signals between Ler <sup>CTD</sup> and 3A3T-DNA. Therefore, our NMR assignments formed a good basis for further DNA binding mechanism studies of Ler.</p></div>\",\"PeriodicalId\":469,\"journal\":{\"name\":\"Applied Magnetic Resonance\",\"volume\":\"54 10\",\"pages\":\"979 - 985\"},\"PeriodicalIF\":1.1000,\"publicationDate\":\"2023-08-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://link.springer.com/content/pdf/10.1007/s00723-023-01575-2.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Applied Magnetic Resonance\",\"FirstCategoryId\":\"101\",\"ListUrlMain\":\"https://link.springer.com/article/10.1007/s00723-023-01575-2\",\"RegionNum\":4,\"RegionCategory\":\"物理与天体物理\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"PHYSICS, ATOMIC, MOLECULAR & CHEMICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Applied Magnetic Resonance","FirstCategoryId":"101","ListUrlMain":"https://link.springer.com/article/10.1007/s00723-023-01575-2","RegionNum":4,"RegionCategory":"物理与天体物理","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"PHYSICS, ATOMIC, MOLECULAR & CHEMICAL","Score":null,"Total":0}
引用次数: 0
摘要
Ler是肠细胞湮没(enterocyte effacement, LEE)致病性岛(pathogenicity island, PAI)基因调控位点的主调控因子,通过抵消H-NS的抑制,激活LEE2到LEE5的转录。Ler包含一个n端寡聚化结构域、一个连接子区域和一个c端DNA结合结构域。然而,Ler的DNA结合机制尚不清楚。在这里,我们报告了LerCTD/3A3T-DNA复合物的1H, 13C和15N NMR分配。我们已经完成了97.8%的LerCTD在配合物中的主链和侧链共振分配,不包括13C '共振。此外,3A3T-DNA约91.5%的1H共振已被分配。我们对该复合物的近乎完整的定位将显著推进Ler CTD和3A3T-DNA之间分子间NOE信号的解析。因此,我们的NMR鉴定为进一步研究Ler的DNA结合机制奠定了良好的基础。
1H, 13C, and 15N Resonance Assignments of the DNA Binding Domain of Ler from Enteropathogenic E. coli in Complex with DNA
Ler is a master regulator for the gene regulation of the locus of enterocyte effacement (LEE) pathogenicity island (PAI), which could activate the transcription of LEE2 to LEE5 by counteracting the repression of H-NS. Ler contains an N-terminal oligomerization domain, a linker region, and a C-terminal DNA binding domain. However, the DNA binding mechanism of Ler remains unclear. Here, we report the 1H, 13C, and 15N NMR assignments of LerCTD/3A3T-DNA complex. We have achieved 97.8% completeness for the backbone and side-chain resonance assignments of LerCTD in the complex, excluding 13C′ resonances. In addition, about 91.5% of the 1H resonances of 3A3T-DNA have been assigned. Our near complete assignments of this complex will significantly advance the parsing of the intermolecular NOE signals between Ler CTD and 3A3T-DNA. Therefore, our NMR assignments formed a good basis for further DNA binding mechanism studies of Ler.
期刊介绍:
Applied Magnetic Resonance provides an international forum for the application of magnetic resonance in physics, chemistry, biology, medicine, geochemistry, ecology, engineering, and related fields.
The contents include articles with a strong emphasis on new applications, and on new experimental methods. Additional features include book reviews and Letters to the Editor.