TY - JOUR
T1 - Novel Twin-Arginine Translocation Pathway-Dependent Phenotypes of Bacillus subtilis Unveiled by Quantitative Proteomics
AU - Goosens, Vivianne J.
AU - Otto, Andreas
AU - Glasner, Corinna
AU - Monteferrante, Carmine C.
AU - van der Ploeg, Rene
AU - Hecker, Michael
AU - Becher, Doerte
AU - van Dijl, Jan Maarten
PY - 2013/2
Y1 - 2013/2
N2 - The twin-arginine translocation (Tat) pathway is known to translocate fully folded proteins across bacterial, archaeal, and organellar membranes. To date, the mechanisms involved in processing, proofreading, and quality control of Tat substrates have remained largely elusive. Bacillus subtilis is an industrially relevant Gram-positive model bacterium. The Tat pathway in B. subtilis differs from that of other well-studied organisms in that it is composed of two complexes operating in parallel. To obtain a better understanding of this pathway in B. subtilis and to identify Tat-associated proteins, the B. subtilis 'Tat proteome' was investigated by quantitative proteomics. Metabolically labeled proteins from cytoplasmic, membrane, and extracellular fractions were analyzed by LC MS/MS. Changes in the amounts of identified peptides allowed for quantitative comparisons of their abundance in tat mutant strains. The observed differences were suggestive of indirect or direct protein protein relationships. The rich data set generated was then approached in hypothesis-driving and hypothesis-driven manners. The hypothesis-driving approach led to the identification of a novel delayed biofilm phenotype of certain tat mutant strains, whereas the hypothesis-driven approach identified the membrane protein QcrA as a new Tat substrate of B. subtilis. Thus, our quantitative proteomics analyses have unveiled novel Tat pathway-dependent phenotypes in Bacillus.
AB - The twin-arginine translocation (Tat) pathway is known to translocate fully folded proteins across bacterial, archaeal, and organellar membranes. To date, the mechanisms involved in processing, proofreading, and quality control of Tat substrates have remained largely elusive. Bacillus subtilis is an industrially relevant Gram-positive model bacterium. The Tat pathway in B. subtilis differs from that of other well-studied organisms in that it is composed of two complexes operating in parallel. To obtain a better understanding of this pathway in B. subtilis and to identify Tat-associated proteins, the B. subtilis 'Tat proteome' was investigated by quantitative proteomics. Metabolically labeled proteins from cytoplasmic, membrane, and extracellular fractions were analyzed by LC MS/MS. Changes in the amounts of identified peptides allowed for quantitative comparisons of their abundance in tat mutant strains. The observed differences were suggestive of indirect or direct protein protein relationships. The rich data set generated was then approached in hypothesis-driving and hypothesis-driven manners. The hypothesis-driving approach led to the identification of a novel delayed biofilm phenotype of certain tat mutant strains, whereas the hypothesis-driven approach identified the membrane protein QcrA as a new Tat substrate of B. subtilis. Thus, our quantitative proteomics analyses have unveiled novel Tat pathway-dependent phenotypes in Bacillus.
KW - Bacillus subtilis
KW - quantitative proteomics
KW - metabolic labeling
KW - twin-arginine translocation
KW - QcrA
KW - biofilm
KW - MEMBRANE-PROTEIN TOPOLOGY
KW - FOLDING QUALITY-CONTROL
KW - GRAM-POSITIVE BACTERIA
KW - RIESKE FE/S PROTEIN
KW - BIOFILM FORMATION
KW - ESCHERICHIA-COLI
KW - SIGNAL PEPTIDES
KW - TAT PATHWAY
KW - TRANSMEMBRANE TOPOLOGY
KW - LEGIONELLA-PNEUMOPHILA
U2 - 10.1021/pr300866f
DO - 10.1021/pr300866f
M3 - Article
SN - 1535-3893
VL - 12
SP - 796
EP - 807
JO - Journal of Proteome Research
JF - Journal of Proteome Research
IS - 2
ER -