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The unphosphorylated form of the PilR two-component system regulates pilA gene expression in Geobacter sulfurreducens

  • 4th International Symposium on Environmental Biotechnology and Engineering-2014
  • Published:
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Abstract

In Geobacter sulfurreducens, metal reduction and generation of bioelectricity require the participation of several elements, and among them, the type IV pili has an essential role. The pilus is composed of multiple PilA monomers. Expression of pilA gene depends mainly on the σ54 factor and the response regulator protein PilR. In this work, we characterized the role of the PilS-PilR two-component system in the regulation of the pilA gene expression. Experimental evidence indicates that PilS is autophosphorylated at the His-334 residue, which in turn is transferred to the conserved Asp-53 in PilR. Contrary to other PilS-PilR systems, substitution D53N in PilR resulted in higher activation of the pilA gene. By using a pilA::luxCDABE fusion with different promoter fragments and in vitro DNA-binding assays, we demonstrated the existence of multiple functional PilR binding sites. A regulatory model in which the non-phosphorylated PilR protein directs activation of pilA expression by binding to two sites in the promoter region of this gene is presented.

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Acknowledgments

This work was supported by CONACyT-179684 grant. AHE was the recipient of a CONACyT postdoctoral fellowship. AA was the recipient of a postdoctoral fellowship from ICyTDF and DGAPA, UNAM. We thank Ana Lilia Tirado Chamu and Paúl Gaytan for technical support and Alejandro Huerta for sharing pCS26-Pac plasmid. Oligonucleotides and automated sequencing was performed at Unit for DNA Sequence and Synthesis of the Instituto de Biotecnología, UNAM.

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Authors and Affiliations

  1. Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001. Col. Chamilpa, Cuernavaca, Morelos, 62210, México

    Alberto Hernández-Eligio, Ángel Andrade, Lizeth Soto, Enrique Morett & Katy Juárez

  2. CONACYT-Research Fellow, Ciudad de México, México

    Alberto Hernández-Eligio

  3. Instituto Nacional de Medicina Genómica, Periférico Sur 4809, Arenal Tepepan, Tlalpan, Ciudad de México, D.F., 14610, México

    Enrique Morett

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  1. Alberto Hernández-Eligio
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  2. Ángel Andrade
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  3. Lizeth Soto
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  4. Enrique Morett
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  5. Katy Juárez
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Correspondence to Katy Juárez.

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Responsible editor: Philippe Garrigues

Alberto Hernández-Eligio CONACyT Research Fellow

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Hernández-Eligio, A., Andrade, Á., Soto, L. et al. The unphosphorylated form of the PilR two-component system regulates pilA gene expression in Geobacter sulfurreducens . Environ Sci Pollut Res 24, 25693–25701 (2017). https://doi.org/10.1007/s11356-016-6192-5

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  • DOI: https://doi.org/10.1007/s11356-016-6192-5

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