The twin-arginine translocation (Tat) systems from Bacillus subtilis display a conserved mode of complex organization and similar substrate recognition requirements

James P Barnett, René van der Ploeg, Robyn T Eijlander, Anja Nenninger, Sharon Mendel, Rense Rozeboom, Oscar P Kuipers, Jan Maarten van Dijl, Colin Robinson

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The twin arginine translocation (Tat) system transports folded proteins across the bacterial plasma membrane. In Gram-negative bacteria, membrane-bound TatABC subunits are all essential for activity, whereas Gram-positive bacteria usually contain only TatAC subunits. In Bacillus subtilis, two TatAC-type systems, TatAdCd and TatAyCy, operate in parallel with different substrate specificities. Here, we show that they recognize similar signal peptide determinants. Both systems translocate green fluorescent protein fused to three distinct Escherichia coli Tat signal peptides, namely DmsA, AmiA and MdoD, and mutagenesis of the DmsA signal peptide confirmed that both Tat pathways recognize similar targeting determinants within Tat signals. Although another E. coli Tat substrate, trimethylamine N-oxide reductase, was translocated by TatAdCd but not by TatAyCy, we conclude that these systems are not predisposed to recognize only specific Tat signal peptides, as suggested by their narrow substrate specificities in B. subtilis. We also analysed complexes involved in the second Tat pathway in B. subtilis, TatAyCy. This revealed a discrete TatAyCy complex together with a separate, homogeneous, approximately 200 kDa TatAy complex. The latter complex differs significantly from the corresponding E. coli TatA complexes, pointing to major structural differences between Tat complexes from Gram-negative and Gram-positive organisms. Like TatAd, TatAy is also detectable in the form of massive cytosolic complexes.

Original languageEnglish
Pages (from-to)232-43
Number of pages12
JournalThe FEBS journal
Volume276
Issue number1
DOIs
Publication statusPublished - Jan 2009

Fingerprint

Bacilli
Bacillus subtilis
Arginine
Protein Sorting Signals
Substrates
Escherichia coli
Substrate Specificity
Bacteria
Mutagenesis
Bacterial Proteins
Gram-Positive Bacteria
Cell membranes
Green Fluorescent Proteins
Twin-Arginine-Translocation System
Gram-Negative Bacteria
Carrier Proteins
Cell Membrane
Membranes

Keywords

  • Adenosine Triphosphate
  • Bacillus subtilis
  • Bacterial Proteins
  • Cell Membrane
  • Conserved Sequence
  • Escherichia coli
  • Gene Products, tat
  • Genes, Reporter
  • Gram-Negative Bacteria
  • Oxidoreductases, N-Demethylating
  • Protein Transport
  • Substrate Specificity
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

The twin-arginine translocation (Tat) systems from Bacillus subtilis display a conserved mode of complex organization and similar substrate recognition requirements. / Barnett, James P; van der Ploeg, René; Eijlander, Robyn T; Nenninger, Anja; Mendel, Sharon; Rozeboom, Rense; Kuipers, Oscar P; van Dijl, Jan Maarten; Robinson, Colin.

In: The FEBS journal, Vol. 276, No. 1, 01.2009, p. 232-43.

Research output: Contribution to journalArticle

Barnett, James P ; van der Ploeg, René ; Eijlander, Robyn T ; Nenninger, Anja ; Mendel, Sharon ; Rozeboom, Rense ; Kuipers, Oscar P ; van Dijl, Jan Maarten ; Robinson, Colin. / The twin-arginine translocation (Tat) systems from Bacillus subtilis display a conserved mode of complex organization and similar substrate recognition requirements. In: The FEBS journal. 2009 ; Vol. 276, No. 1. pp. 232-43.
@article{d39c04cfe2d847b1a9864498ec4101b3,
title = "The twin-arginine translocation (Tat) systems from Bacillus subtilis display a conserved mode of complex organization and similar substrate recognition requirements",
abstract = "The twin arginine translocation (Tat) system transports folded proteins across the bacterial plasma membrane. In Gram-negative bacteria, membrane-bound TatABC subunits are all essential for activity, whereas Gram-positive bacteria usually contain only TatAC subunits. In Bacillus subtilis, two TatAC-type systems, TatAdCd and TatAyCy, operate in parallel with different substrate specificities. Here, we show that they recognize similar signal peptide determinants. Both systems translocate green fluorescent protein fused to three distinct Escherichia coli Tat signal peptides, namely DmsA, AmiA and MdoD, and mutagenesis of the DmsA signal peptide confirmed that both Tat pathways recognize similar targeting determinants within Tat signals. Although another E. coli Tat substrate, trimethylamine N-oxide reductase, was translocated by TatAdCd but not by TatAyCy, we conclude that these systems are not predisposed to recognize only specific Tat signal peptides, as suggested by their narrow substrate specificities in B. subtilis. We also analysed complexes involved in the second Tat pathway in B. subtilis, TatAyCy. This revealed a discrete TatAyCy complex together with a separate, homogeneous, approximately 200 kDa TatAy complex. The latter complex differs significantly from the corresponding E. coli TatA complexes, pointing to major structural differences between Tat complexes from Gram-negative and Gram-positive organisms. Like TatAd, TatAy is also detectable in the form of massive cytosolic complexes.",
keywords = "Adenosine Triphosphate, Bacillus subtilis, Bacterial Proteins, Cell Membrane, Conserved Sequence, Escherichia coli, Gene Products, tat, Genes, Reporter, Gram-Negative Bacteria, Oxidoreductases, N-Demethylating, Protein Transport, Substrate Specificity, Journal Article, Research Support, Non-U.S. Gov't",
author = "Barnett, {James P} and {van der Ploeg}, Ren{\'e} and Eijlander, {Robyn T} and Anja Nenninger and Sharon Mendel and Rense Rozeboom and Kuipers, {Oscar P} and {van Dijl}, {Jan Maarten} and Colin Robinson",
year = "2009",
month = "1",
doi = "10.1111/j.1742-4658.2008.06776.x",
language = "English",
volume = "276",
pages = "232--43",
journal = "FEBS Journal",
issn = "1742-464X",
publisher = "Wiley",
number = "1",

}

TY - JOUR

T1 - The twin-arginine translocation (Tat) systems from Bacillus subtilis display a conserved mode of complex organization and similar substrate recognition requirements

AU - Barnett, James P

AU - van der Ploeg, René

AU - Eijlander, Robyn T

AU - Nenninger, Anja

AU - Mendel, Sharon

AU - Rozeboom, Rense

AU - Kuipers, Oscar P

AU - van Dijl, Jan Maarten

AU - Robinson, Colin

PY - 2009/1

Y1 - 2009/1

N2 - The twin arginine translocation (Tat) system transports folded proteins across the bacterial plasma membrane. In Gram-negative bacteria, membrane-bound TatABC subunits are all essential for activity, whereas Gram-positive bacteria usually contain only TatAC subunits. In Bacillus subtilis, two TatAC-type systems, TatAdCd and TatAyCy, operate in parallel with different substrate specificities. Here, we show that they recognize similar signal peptide determinants. Both systems translocate green fluorescent protein fused to three distinct Escherichia coli Tat signal peptides, namely DmsA, AmiA and MdoD, and mutagenesis of the DmsA signal peptide confirmed that both Tat pathways recognize similar targeting determinants within Tat signals. Although another E. coli Tat substrate, trimethylamine N-oxide reductase, was translocated by TatAdCd but not by TatAyCy, we conclude that these systems are not predisposed to recognize only specific Tat signal peptides, as suggested by their narrow substrate specificities in B. subtilis. We also analysed complexes involved in the second Tat pathway in B. subtilis, TatAyCy. This revealed a discrete TatAyCy complex together with a separate, homogeneous, approximately 200 kDa TatAy complex. The latter complex differs significantly from the corresponding E. coli TatA complexes, pointing to major structural differences between Tat complexes from Gram-negative and Gram-positive organisms. Like TatAd, TatAy is also detectable in the form of massive cytosolic complexes.

AB - The twin arginine translocation (Tat) system transports folded proteins across the bacterial plasma membrane. In Gram-negative bacteria, membrane-bound TatABC subunits are all essential for activity, whereas Gram-positive bacteria usually contain only TatAC subunits. In Bacillus subtilis, two TatAC-type systems, TatAdCd and TatAyCy, operate in parallel with different substrate specificities. Here, we show that they recognize similar signal peptide determinants. Both systems translocate green fluorescent protein fused to three distinct Escherichia coli Tat signal peptides, namely DmsA, AmiA and MdoD, and mutagenesis of the DmsA signal peptide confirmed that both Tat pathways recognize similar targeting determinants within Tat signals. Although another E. coli Tat substrate, trimethylamine N-oxide reductase, was translocated by TatAdCd but not by TatAyCy, we conclude that these systems are not predisposed to recognize only specific Tat signal peptides, as suggested by their narrow substrate specificities in B. subtilis. We also analysed complexes involved in the second Tat pathway in B. subtilis, TatAyCy. This revealed a discrete TatAyCy complex together with a separate, homogeneous, approximately 200 kDa TatAy complex. The latter complex differs significantly from the corresponding E. coli TatA complexes, pointing to major structural differences between Tat complexes from Gram-negative and Gram-positive organisms. Like TatAd, TatAy is also detectable in the form of massive cytosolic complexes.

KW - Adenosine Triphosphate

KW - Bacillus subtilis

KW - Bacterial Proteins

KW - Cell Membrane

KW - Conserved Sequence

KW - Escherichia coli

KW - Gene Products, tat

KW - Genes, Reporter

KW - Gram-Negative Bacteria

KW - Oxidoreductases, N-Demethylating

KW - Protein Transport

KW - Substrate Specificity

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1111/j.1742-4658.2008.06776.x

DO - 10.1111/j.1742-4658.2008.06776.x

M3 - Article

VL - 276

SP - 232

EP - 243

JO - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

IS - 1

ER -