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1H, 13C, and 15N Resonance Assignments of the DNA Binding Domain of Ler from Enteropathogenic E. coli in Complex with DNA

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Abstract

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.

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Availability of Data and Material

The 1H, 13C, and 15N chemical shifts were deposited in the BioMagResBank (https://www.bmrb.wisc.edu.) with BMRB accession number 51961. All other data generated or analyzed during this study are included in this published article.

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Acknowledgements

NMR experiments were performed at the Beijing NMR Center of National Center for Protein Sciences, at Peking University. This work was supported by grant 2016YFA0501200 from Ministry of Science and Technology of China. We thank Dr. Hongwei Li and Dr. Xiaogang Niu for their assistance in the NMR experiments.

Funding

This work was supported by Grant 2016YFA0501200 from Ministry of Science and Technology of China.

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Authors and Affiliations

  1. Beijing Nuclear Magnetic Resonance Center, School of Life Sciences, College of Chemistry and Molecular Engineering, Peking University, Beijing, China

    Fan Liu & Bin Xia

  2. Suzhou Key Laboratory of Pathogen Bioscience and Anti-infective Medicine, MOE Key Laboratory of Geriatric Diseases and Immunology, Institute of Molecular Enzymology & School of Biology and Basic Medical Sciences, Soochow University, Suzhou, China

    Bo Duan

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  1. Fan Liu
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  2. Bo Duan
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Contributions

FL and BD carried out the experiment and analyzed data. FL and BX wrote the manuscript. BX supervised the project.

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Correspondence to Bo Duan or Bin Xia.

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Liu, F., Duan, B. & Xia, B. 1H, 13C, and 15N Resonance Assignments of the DNA Binding Domain of Ler from Enteropathogenic E. coli in Complex with DNA. Appl Magn Reson 54, 979–985 (2023). https://doi.org/10.1007/s00723-023-01575-2

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