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Journal of Bacteriology, July 1999, p. 4004-4011, Vol. 181, No. 13
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Membrane Association and Multimerization of Secreton Component PulC

Odile M. Possot, Manon Gérard-Vincent,dagger and Anthony P. Pugsley*

Unité de Génétique Moléculaire, CNRS URA1773, Institut Pasteur, 75724 Paris Cedex 15, France

Received 15 January 1999/Accepted 17 March 1999

The PulC component of the Klebsiella oxytoca pullulanase secretion machinery (the secreton) was found by subcellular fractionation to be associated with both the cytoplasmic (inner) and outer membranes. Association with the outer membrane was independent of other secreton components, including the outer membrane protein PulD (secretin). The association of PulC with the inner membrane is mediated by the signal anchor sequence located close to its N terminus. These results suggest that PulC forms a bridge between the two membranes that is disrupted when bacteria are broken open for fractionation. Neither the signal anchor sequence nor the cytoplasmic N-terminal region that precedes it was found to be required for PulC function, indicating that PulC does not undergo sequence-specific interactions with other cytoplasmic membrane proteins. Cross-linking of whole cells resulted in the formation of a ca. 110-kDa band that reacted with PulC-specific serum and whose detection depended on the presence of PulD. However, antibodies against PulD failed to react with this band, suggesting that it could be a homo-PulC trimer whose formation requires PulD. The data are discussed in terms of the possible role of PulC in energy transduction for exoprotein secretion.

* Corresponding author. Mailing address: Unité de Génétique Moléculaire, Institut Pasteur, 25, rue du Dr. Roux, 75724 Paris Cedex 15, France. Phone: 33/0-145688494. Fax: 33/0-145688960. E-mail: max{at}pasteur.fr.

dagger Present address: Laboratoire d'Ingénierie et Dynamique des Systèmes Macromoléculaires, IBSM/CNRS, 13402 Marseille Cedex 20, France.

Journal of Bacteriology, July 1999, p. 4004-4011, Vol. 181, No. 13
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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