The chaperone ClpX stimulates expression of Staphylococcus aureus protein A by rot dependent and independent pathways

Lotte Jelsbak; Hanne Ingmer; Lukás Valihrach; Marianne Thorup Cohn; Mie H.G. Christiansen; Birgitte H. Kallipolitis; Dorte Frees

doi:10.1371/journal.pone.0012752

The chaperone ClpX stimulates expression of Staphylococcus aureus protein A by rot dependent and independent pathways

Lotte Jelsbak, Hanne Ingmer, Lukás Valihrach, Marianne Thorup Cohn, Mie H.G. Christiansen, Birgitte H. Kallipolitis, Dorte Frees

27 Citations (Scopus)

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Abstract

The Clp ATPases (Hsp100) constitute a family of closely related proteins that have protein reactivating and remodelling activities typical of molecular chaperones. In Staphylococcus aureus the ClpX chaperone is essential for virulence and for transcription of spa encoding Protein A. The present study was undertaken to elucidate the mechanism by which ClpX stimulates expression of Protein A. For this purpose, we prepared antibodies directed against Rot, an activator of spa transcription, and demonstrated that cells devoid of ClpX contain three-fold less Rot than wild-type cells. By varying Rot expression from an inducible promoter we showed that expression of Protein A requires a threshold level of Rot. In the absence of ClpX the Rot content is reduced below this threshold level, hence, explaining the substantially reduced Protein A expression in the clpX mutant. Experiments addressed at pinpointing the role of ClpX in Rot synthesis revealed that ClpX is required for translation of Rot. Interestingly, translation of the spa mRNA was, like the rot mRNA, enhanced by ClpX. These data demonstrate that ClpX performs dual roles in regulating Protein A expression, as ClpX stimulates transcription of spa by enhancing translation of Rot, and that ClpX additionally is required for full translation of the spa mRNA. The current findings emphasize that ClpX has a central role in fine-tuning virulence regulation in S. aureus.

Original language	English
Journal	PLoS ONE
Volume	5
Issue number	9
Number of pages	11
ISSN	1932-6203
DOIs	https://doi.org/10.1371/journal.pone.0012752
Publication status	Published - 2010

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title = "The chaperone ClpX stimulates expression of Staphylococcus aureus protein A by rot dependent and independent pathways",

abstract = "The Clp ATPases (Hsp100) constitute a family of closely related proteins that have protein reactivating and remodelling activities typical of molecular chaperones. In Staphylococcus aureus the ClpX chaperone is essential for virulence and for transcription of spa encoding Protein A. The present study was undertaken to elucidate the mechanism by which ClpX stimulates expression of Protein A. For this purpose, we prepared antibodies directed against Rot, an activator of spa transcription, and demonstrated that cells devoid of ClpX contain three-fold less Rot than wild-type cells. By varying Rot expression from an inducible promoter we showed that expression of Protein A requires a threshold level of Rot. In the absence of ClpX the Rot content is reduced below this threshold level, hence, explaining the substantially reduced Protein A expression in the clpX mutant. Experiments addressed at pinpointing the role of ClpX in Rot synthesis revealed that ClpX is required for translation of Rot. Interestingly, translation of the spa mRNA was, like the rot mRNA, enhanced by ClpX. These data demonstrate that ClpX performs dual roles in regulating Protein A expression, as ClpX stimulates transcription of spa by enhancing translation of Rot, and that ClpX additionally is required for full translation of the spa mRNA. The current findings emphasize that ClpX has a central role in fine-tuning virulence regulation in S. aureus.",

author = "Lotte Jelsbak and Hanne Ingmer and Luk{\'a}s Valihrach and Cohn, {Marianne Thorup} and Christiansen, {Mie H.G.} and Kallipolitis, {Birgitte H.} and Dorte Frees",

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T1 - The chaperone ClpX stimulates expression of Staphylococcus aureus protein A by rot dependent and independent pathways

AU - Jelsbak, Lotte

AU - Ingmer, Hanne

AU - Valihrach, Lukás

AU - Cohn, Marianne Thorup

AU - Christiansen, Mie H.G.

AU - Kallipolitis, Birgitte H.

AU - Frees, Dorte

N1 - Paper id:: e12752

PY - 2010

Y1 - 2010

N2 - The Clp ATPases (Hsp100) constitute a family of closely related proteins that have protein reactivating and remodelling activities typical of molecular chaperones. In Staphylococcus aureus the ClpX chaperone is essential for virulence and for transcription of spa encoding Protein A. The present study was undertaken to elucidate the mechanism by which ClpX stimulates expression of Protein A. For this purpose, we prepared antibodies directed against Rot, an activator of spa transcription, and demonstrated that cells devoid of ClpX contain three-fold less Rot than wild-type cells. By varying Rot expression from an inducible promoter we showed that expression of Protein A requires a threshold level of Rot. In the absence of ClpX the Rot content is reduced below this threshold level, hence, explaining the substantially reduced Protein A expression in the clpX mutant. Experiments addressed at pinpointing the role of ClpX in Rot synthesis revealed that ClpX is required for translation of Rot. Interestingly, translation of the spa mRNA was, like the rot mRNA, enhanced by ClpX. These data demonstrate that ClpX performs dual roles in regulating Protein A expression, as ClpX stimulates transcription of spa by enhancing translation of Rot, and that ClpX additionally is required for full translation of the spa mRNA. The current findings emphasize that ClpX has a central role in fine-tuning virulence regulation in S. aureus.

AB - The Clp ATPases (Hsp100) constitute a family of closely related proteins that have protein reactivating and remodelling activities typical of molecular chaperones. In Staphylococcus aureus the ClpX chaperone is essential for virulence and for transcription of spa encoding Protein A. The present study was undertaken to elucidate the mechanism by which ClpX stimulates expression of Protein A. For this purpose, we prepared antibodies directed against Rot, an activator of spa transcription, and demonstrated that cells devoid of ClpX contain three-fold less Rot than wild-type cells. By varying Rot expression from an inducible promoter we showed that expression of Protein A requires a threshold level of Rot. In the absence of ClpX the Rot content is reduced below this threshold level, hence, explaining the substantially reduced Protein A expression in the clpX mutant. Experiments addressed at pinpointing the role of ClpX in Rot synthesis revealed that ClpX is required for translation of Rot. Interestingly, translation of the spa mRNA was, like the rot mRNA, enhanced by ClpX. These data demonstrate that ClpX performs dual roles in regulating Protein A expression, as ClpX stimulates transcription of spa by enhancing translation of Rot, and that ClpX additionally is required for full translation of the spa mRNA. The current findings emphasize that ClpX has a central role in fine-tuning virulence regulation in S. aureus.

U2 - 10.1371/journal.pone.0012752

DO - 10.1371/journal.pone.0012752

M3 - Journal article

SN - 1932-6203

VL - 5

JO - PLoS ONE

JF - PLoS ONE

IS - 9

ER -

The chaperone ClpX stimulates expression of Staphylococcus aureus protein A by rot dependent and independent pathways

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