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The Haemophilus influenzae Hia autotransporter contains an unusually short trimeric translocator domain.

Publication ,  Journal Article
Surana, NK; Cutter, D; Barenkamp, SJ; St Geme, JW
Published in: J Biol Chem
April 9, 2004

Gram-negative bacterial autotransporter proteins are a growing group of virulence factors that are characterized by their ability to cross the outer membrane without the help of accessory proteins. A conserved C-terminal beta-domain is critical for targeting of autotransporters to the outer membrane and for translocation of the N-terminal "passenger" domain to the bacterial surface. We have demonstrated previously that the Haemophilus influenzae Hia adhesin belongs to the autotransporter family, with translocator activity residing in the C-terminal 319 residues. To gain further insight into the mechanism of autotransporter protein translocation, we performed a structure-function analysis on Hia. In initial experiments, we generated a series of in-frame deletions and a set of chimeric proteins containing varying regions of the Hia C terminus fused to a heterologous passenger domain and discovered that the final 76 residues of Hia are both necessary and sufficient for translocation. Analysis by flow cytometry revealed that the region N-terminal to this shortened translocator domain is surface localized, further suggesting that this region is not involved in beta-barrel formation or in translocation of the passenger domain. Western analysis demonstrated that the translocation-competent regions of the C terminus migrated at masses consistent with trimers, suggesting that the Hia C terminus oligomerizes. Furthermore, fusion proteins containing a heterologous passenger domain demonstrated that similarly small C-terminal regions of Yersinia sp. YadA and Neisseria meningitidis NhhA are translocation-competent. These data provide experimental support for a unique subclass of autotransporters characterized by a short trimeric translocator domain.

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Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

April 9, 2004

Volume

279

Issue

15

Start / End Page

14679 / 14685

Location

United States

Related Subject Headings

  • Yersinia
  • Subcellular Fractions
  • Structure-Activity Relationship
  • Sequence Homology, Amino Acid
  • Recombinant Fusion Proteins
  • Protein Transport
  • Protein Structure, Tertiary
  • Protein Structure, Secondary
  • Plasmids
  • Neisseria meningitidis
 

Citation

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Surana, N. K., Cutter, D., Barenkamp, S. J., & St Geme, J. W. (2004). The Haemophilus influenzae Hia autotransporter contains an unusually short trimeric translocator domain. J Biol Chem, 279(15), 14679–14685. https://doi.org/10.1074/jbc.M311496200
Surana, Neeraj K., David Cutter, Stephen J. Barenkamp, and Joseph W. St Geme. “The Haemophilus influenzae Hia autotransporter contains an unusually short trimeric translocator domain.J Biol Chem 279, no. 15 (April 9, 2004): 14679–85. https://doi.org/10.1074/jbc.M311496200.
Surana NK, Cutter D, Barenkamp SJ, St Geme JW. The Haemophilus influenzae Hia autotransporter contains an unusually short trimeric translocator domain. J Biol Chem. 2004 Apr 9;279(15):14679–85.
Surana, Neeraj K., et al. “The Haemophilus influenzae Hia autotransporter contains an unusually short trimeric translocator domain.J Biol Chem, vol. 279, no. 15, Apr. 2004, pp. 14679–85. Pubmed, doi:10.1074/jbc.M311496200.
Surana NK, Cutter D, Barenkamp SJ, St Geme JW. The Haemophilus influenzae Hia autotransporter contains an unusually short trimeric translocator domain. J Biol Chem. 2004 Apr 9;279(15):14679–14685.

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

April 9, 2004

Volume

279

Issue

15

Start / End Page

14679 / 14685

Location

United States

Related Subject Headings

  • Yersinia
  • Subcellular Fractions
  • Structure-Activity Relationship
  • Sequence Homology, Amino Acid
  • Recombinant Fusion Proteins
  • Protein Transport
  • Protein Structure, Tertiary
  • Protein Structure, Secondary
  • Plasmids
  • Neisseria meningitidis