The impact of a His-tag on DNA binding by RNA polymerase alpha-C-terminal domain from Helicobacter pylori

In this study, we demonstrate that a His-tag impacts DNA complex formation by the C-terminal domain of the ??-subunit (??CTD) of Helicobacter pylori RNA polymerase in a metal-dependent fashion. The ??CTD was purified with a cleavable His-tag, and complex formation between αCTD, the nickel-responsive metalloregulator HpNikR, and DNA was investigated using electrophoretic mobility shift assays. An interaction between His-tagged ??CTD (His??CTD) and the HpNikR-DNA complex was observed; however, this interaction was not observed upon removal of the His-tag. Further analysis revealed that complex formation between HisαCTD and DNA is non-specific and dependent on the type of metal ions present. Overall, the results indicate that a histidine tag is able to modulate DNA-binding activity and suggests that the impact of metal affinity tags should be considered when analyzing the in vitro biomolecular interactions of metalloproteins.
Source: Protein Expression and Purification - Category: Biochemistry Source Type: research

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In the original publication of the article, the figure  3 was published with errors. The corrected figure 3 should appear as in this correction.
Source: Journal of Gastroenterology - Category: Gastroenterology Source Type: research
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Source: RSC - Chem. Commun. latest articles - Category: Chemistry Authors: Source Type: research
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Source: Cochrane News and Events - Category: Information Technology Authors: Source Type: news
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Source: Chemical Communications - Category: Chemistry Authors: Tags: Chem Commun (Camb) Source Type: research
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Source: Oncology Letters - Category: Cancer & Oncology Tags: Oncol Lett Source Type: research
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Source: Indian Journal of Gastroenterology - Category: Gastroenterology Source Type: research
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Source: Cellular and Molecular Immunology - Category: Molecular Biology Authors: Tags: Cell Mol Immunol Source Type: research
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Source: European Journal of Cancer Prevention - Category: Cancer & Oncology Tags: Letters to the Editor Source Type: research
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Source: Cell Communication and Signaling - Category: Molecular Biology Authors: Tags: Research Source Type: research
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