The metalloprotease SepA governs processing of accumulation-associated protein and shapes intercellular adhesive surface properties in Staphylococcus epidermidis

Alexandra E. Paharik, Marta Kotasinska, Anna Both, Tra My N Hoang, Henning Büttner, Paroma Roy, Paul D Fey, Alexander R. Horswill, Holger Rohde

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The otherwise harmless skin inhabitant Staphylococcus epidermidis is a major cause of healthcare-associated medical device infections. The species' selective pathogenic potential depends on its production of surface adherent biofilms. The Cell wall-anchored protein Aap promotes biofilm formation in S. epidermidis, independently from the polysaccharide intercellular adhesin PIA. Aap requires proteolytic cleavage to act as an intercellular adhesin. Whether and which staphylococcal proteases account for Aap processing is yet unknown. Here, evidence is provided that in PIA-negative S. epidermidis 1457Δica, the metalloprotease SepA is required for Aap-dependent S. epidermidis biofilm formation in static and dynamic biofilm models. qRT-PCR and protease activity assays demonstrated that under standard growth conditions, sepA is repressed by the global regulator SarA. Inactivation of sarA increased SepA production, and in turn augmented biofilm formation. Genetic and biochemical analyses demonstrated that SepA-related induction of biofilm accumulation resulted from enhanced Aap processing. Studies using recombinant proteins demonstrated that SepA is able to cleave the A domain of Aap at residue 335 and between the A and B domains at residue 601. This study identifies the mechanism behind Aap-mediated biofilm maturation, and also demonstrates a novel role for a secreted staphylococcal protease as a requirement for the development of a biofilm.

Original languageEnglish (US)
Pages (from-to)860-874
Number of pages15
JournalMolecular Microbiology
Volume103
Issue number5
DOIs
StatePublished - Mar 1 2017

Fingerprint

Staphylococcus epidermidis
Surface Properties
Metalloproteases
Biofilms
Adhesives
Proteins
Recombinant Proteins
Cell Wall
Molecular Biology
Peptide Hydrolases
Delivery of Health Care
Equipment and Supplies
Polymerase Chain Reaction
Skin
Growth
Infection

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

The metalloprotease SepA governs processing of accumulation-associated protein and shapes intercellular adhesive surface properties in Staphylococcus epidermidis. / Paharik, Alexandra E.; Kotasinska, Marta; Both, Anna; Hoang, Tra My N; Büttner, Henning; Roy, Paroma; Fey, Paul D; Horswill, Alexander R.; Rohde, Holger.

In: Molecular Microbiology, Vol. 103, No. 5, 01.03.2017, p. 860-874.

Research output: Contribution to journalArticle

Paharik, Alexandra E. ; Kotasinska, Marta ; Both, Anna ; Hoang, Tra My N ; Büttner, Henning ; Roy, Paroma ; Fey, Paul D ; Horswill, Alexander R. ; Rohde, Holger. / The metalloprotease SepA governs processing of accumulation-associated protein and shapes intercellular adhesive surface properties in Staphylococcus epidermidis. In: Molecular Microbiology. 2017 ; Vol. 103, No. 5. pp. 860-874.
@article{6ab0df216a414520b40eefa89c72ab77,
title = "The metalloprotease SepA governs processing of accumulation-associated protein and shapes intercellular adhesive surface properties in Staphylococcus epidermidis",
abstract = "The otherwise harmless skin inhabitant Staphylococcus epidermidis is a major cause of healthcare-associated medical device infections. The species' selective pathogenic potential depends on its production of surface adherent biofilms. The Cell wall-anchored protein Aap promotes biofilm formation in S. epidermidis, independently from the polysaccharide intercellular adhesin PIA. Aap requires proteolytic cleavage to act as an intercellular adhesin. Whether and which staphylococcal proteases account for Aap processing is yet unknown. Here, evidence is provided that in PIA-negative S. epidermidis 1457Δica, the metalloprotease SepA is required for Aap-dependent S. epidermidis biofilm formation in static and dynamic biofilm models. qRT-PCR and protease activity assays demonstrated that under standard growth conditions, sepA is repressed by the global regulator SarA. Inactivation of sarA increased SepA production, and in turn augmented biofilm formation. Genetic and biochemical analyses demonstrated that SepA-related induction of biofilm accumulation resulted from enhanced Aap processing. Studies using recombinant proteins demonstrated that SepA is able to cleave the A domain of Aap at residue 335 and between the A and B domains at residue 601. This study identifies the mechanism behind Aap-mediated biofilm maturation, and also demonstrates a novel role for a secreted staphylococcal protease as a requirement for the development of a biofilm.",
author = "Paharik, {Alexandra E.} and Marta Kotasinska and Anna Both and Hoang, {Tra My N} and Henning B{\"u}ttner and Paroma Roy and Fey, {Paul D} and Horswill, {Alexander R.} and Holger Rohde",
year = "2017",
month = "3",
day = "1",
doi = "10.1111/mmi.13594",
language = "English (US)",
volume = "103",
pages = "860--874",
journal = "Molecular Microbiology",
issn = "0950-382X",
publisher = "Wiley-Blackwell",
number = "5",

}

TY - JOUR

T1 - The metalloprotease SepA governs processing of accumulation-associated protein and shapes intercellular adhesive surface properties in Staphylococcus epidermidis

AU - Paharik, Alexandra E.

AU - Kotasinska, Marta

AU - Both, Anna

AU - Hoang, Tra My N

AU - Büttner, Henning

AU - Roy, Paroma

AU - Fey, Paul D

AU - Horswill, Alexander R.

AU - Rohde, Holger

PY - 2017/3/1

Y1 - 2017/3/1

N2 - The otherwise harmless skin inhabitant Staphylococcus epidermidis is a major cause of healthcare-associated medical device infections. The species' selective pathogenic potential depends on its production of surface adherent biofilms. The Cell wall-anchored protein Aap promotes biofilm formation in S. epidermidis, independently from the polysaccharide intercellular adhesin PIA. Aap requires proteolytic cleavage to act as an intercellular adhesin. Whether and which staphylococcal proteases account for Aap processing is yet unknown. Here, evidence is provided that in PIA-negative S. epidermidis 1457Δica, the metalloprotease SepA is required for Aap-dependent S. epidermidis biofilm formation in static and dynamic biofilm models. qRT-PCR and protease activity assays demonstrated that under standard growth conditions, sepA is repressed by the global regulator SarA. Inactivation of sarA increased SepA production, and in turn augmented biofilm formation. Genetic and biochemical analyses demonstrated that SepA-related induction of biofilm accumulation resulted from enhanced Aap processing. Studies using recombinant proteins demonstrated that SepA is able to cleave the A domain of Aap at residue 335 and between the A and B domains at residue 601. This study identifies the mechanism behind Aap-mediated biofilm maturation, and also demonstrates a novel role for a secreted staphylococcal protease as a requirement for the development of a biofilm.

AB - The otherwise harmless skin inhabitant Staphylococcus epidermidis is a major cause of healthcare-associated medical device infections. The species' selective pathogenic potential depends on its production of surface adherent biofilms. The Cell wall-anchored protein Aap promotes biofilm formation in S. epidermidis, independently from the polysaccharide intercellular adhesin PIA. Aap requires proteolytic cleavage to act as an intercellular adhesin. Whether and which staphylococcal proteases account for Aap processing is yet unknown. Here, evidence is provided that in PIA-negative S. epidermidis 1457Δica, the metalloprotease SepA is required for Aap-dependent S. epidermidis biofilm formation in static and dynamic biofilm models. qRT-PCR and protease activity assays demonstrated that under standard growth conditions, sepA is repressed by the global regulator SarA. Inactivation of sarA increased SepA production, and in turn augmented biofilm formation. Genetic and biochemical analyses demonstrated that SepA-related induction of biofilm accumulation resulted from enhanced Aap processing. Studies using recombinant proteins demonstrated that SepA is able to cleave the A domain of Aap at residue 335 and between the A and B domains at residue 601. This study identifies the mechanism behind Aap-mediated biofilm maturation, and also demonstrates a novel role for a secreted staphylococcal protease as a requirement for the development of a biofilm.

UR - http://www.scopus.com/inward/record.url?scp=85013748598&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85013748598&partnerID=8YFLogxK

U2 - 10.1111/mmi.13594

DO - 10.1111/mmi.13594

M3 - Article

C2 - 27997732

AN - SCOPUS:85013748598

VL - 103

SP - 860

EP - 874

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

IS - 5

ER -