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The transmembrane domain of the DnaJ-like protein DjlA is a dimerisation domain

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Abstract

DjlA is a bitopic inner membrane protein, which belongs to the DnaJ co-chaperone family in Escherichia coli. Overproduction of DjlA leads to the synthesis of colanic acid, resulting in mucoidy, via the activation of the two-component regulatory system RcsC/B that controls the cps (capsular polysaccharide) operon. This induction requires both the co-chaperone activity of DjlA, in cooperation with DnaK and GrpE, and its unique transmembrane (TM) domain. Here, we show that the TM segment of DjlA acts as a dimerisation domain: when fused to the N-terminal DNA-binding domain of the lambda cI repressor protein, it can substitute for the native C-terminal dimerisation domain of cI, thus generating an active cI repressor. Replacing the TM domain of DjlA by other TM domains, with or without dimerising capacity, revealed that dimerisation is not sufficient for the induction of cps expression, indicating an additional sequence- or structurally specific role for the TM domain. Finally, the conserved glycines present in the TM domain of DjlA are essential for the induction of mucoidy, but not for dimerisation.

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Acknowledgements

This work was supported by the French Ministry of Research (grant "PRFMMIP" to I.B.H and A.J.; doctoral fellowship to C.M.T.) and by the National Institutes of Health (grant GM18569-01 to J.A.L and grant GM54160-02 to J.B). D.J.C. was the recipient of an International Traveling Fellowship from the Wellcome Trust (Ref. No. 043080/Z/94/Z) and a postdoctoral fellowship from the Fondation de la Recherche Médicale. J.B. is an American Cancer Society Research Professor. We are thankful to Marc Tempête for expert technical assistance

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Author notes

  1. D. J. Clarke

    Present address: Department of Biology and Biochemistry, University of Bath, Bath, BA2 7A, UK

  2. J. A. Leeds

    Present address: Dyax Corporation, Cambridge, MA 02139, USA

Authors and Affiliations

  1. Institut de Génétique et Microbiologie, UMR CNRS 8621, Bâtiment 400, Université Paris-Sud, 91405 , Orsay, France

    C. M. Toutain, D. J. Clarke, J. Kuhn, I. B. Holland & A. Jacq

  2. Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA

    J. A. Leeds & J. Beckwith

Authors
  1. C. M. Toutain
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  2. D. J. Clarke
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  3. J. A. Leeds
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  4. J. Kuhn
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  5. J. Beckwith
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  6. I. B. Holland
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  7. A. Jacq
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Correspondence to A. Jacq.

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Communicated by R. Devoret

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Toutain, C.M., Clarke, D.J., Leeds, J.A. et al. The transmembrane domain of the DnaJ-like protein DjlA is a dimerisation domain. Mol Gen Genomics 268, 761–770 (2003). https://doi.org/10.1007/s00438-002-0793-z

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  • DOI: https://doi.org/10.1007/s00438-002-0793-z

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