Archives of Microbiology

, Volume 172, Issue 4, pp 227–232

The role of the twin-arginine motif in the signal peptide encoded by the hydA gene of the hydrogenase from Wolinella succinogenes

  • Roland Gross
  • Jörg Simon
  • A. Kröger
Original paper

DOI: 10.1007/s002030050764

Cite this article as:
Gross, R., Simon, J. & Kröger, A. Arch Microbiol (1999) 172: 227. doi:10.1007/s002030050764

Abstract

The hydABC operon of Wolinella succinogenes encodes the three subunits of the membrane-integrated Ni-hydrogenase. The catalytic subunit, HydB, is on the periplasmic side of the membrane. Residues R41 and R42 of the twin-arginine motif within the signal peptide of the precursor of the iron-sulfur subunit, HydA, were replaced by two glutamine residues. The corresponding mutant did not grow with H2 as the electron donor of anaerobic respiration. Mature HydB and the precursor protein of HydA were located exclusively in the cytoplasmic cell fraction of the mutant, which catalyzed the reduction of benzyl viologen by H2, suggesting that HydB contained Ni. The HydC protein was located in the membrane fraction of the mutant in wild-type amounts. HydC was purified and was shown to contain heme. The results suggest that HydA and HydB are translocated across the membrane by the Tat (twin-arginine translocation) system. The translocation of HydA and HydB as well as the maturation of the precursor protein of HydA appear to depend on the presence of the twin-arginine motif. In contrast, maturation of HydB, the insertion of HydC into the membrane, and heme attachment to HydC are apparently independent of the twin-arginine motif and do not require translocation of the two other hydrogenase subunits.

Key words Ni-hydrogenaseCytochrome bWolinella succinogenesSignal peptideTwin-arginine transferTat system

Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • Roland Gross
    • 1
  • Jörg Simon
    • 1
  • A. Kröger
    • 1
  1. 1.Institut für Mikrobiologie, Johann Wolfgang Goethe-Universität, Marie-Curie-Straße 9, D-60439 Frankfurt am Main, Germany e-mail: a.kroeger@em.uni-frankfurt.de, Tel.: +49-69-79829507, Fax: +49-69-79829527DE