The sequences of MinE responsible for its subcellular localization analyzed by competitive binding method in Escherichia coli
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The subcellular localization of a protein is important for its proper function. Escherichia coli MinE is a small protein with clear subcellular localization, which provides a good model to study protein localization mechanism. In the present study, a series of recombinant minEs truncated in one end or in the middle regions, fused with egfp, was constructed, and these recombinant proteins could compete to function with the chromosomal MinE. Our results showed that the sequences related to the subcellular localization of MinE span several functional domains, demonstrating that MinE positioning in cells depends on multiple factors. The eGFP fusions with some truncated MinE from N-terminal resulted in different cell phenotypes and localization features, implying that these fusions can interfere chromosomal MinE’s function, similar to MinE36–88 phenotype in the previous report. The amino acid in the region (32–48) is sensitive to change MinE conformation and influence its dimerization. Some truncated protein structure could be unstable. Thus, the MinE localization is prerequisite for its proper anti-MinCD function and some new features of MinE were demonstrated. This approach can be extended for subcellular localization research for other essential proteins.
KeywordsMinE Subcellular localization Cell division Sequence
This work was supported by the Consejo Nacional de Ciencia y Tecnología (CONACyT)-México (Grant No. 168541) and Secretaría de Investigación y Posgrado del Instituto Politécnico Nacional, México (No. 20151373). Miguel Ángel Pérez-Rodríguez held scholarships from CONACyT. X. Guo and Mario A. Rodriguez Perez hold scholarships from COFAA-IPN.
Compliance with ethical standards
The authors declare that they have no conflict of interest.
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