A Novel Repressor of the <italic toggle="yes">ica</italic> Locus Discovered in Clinically Isolated Super-Biofilm-Elaborating <italic toggle="yes">Staphylococcus aureus</italic>

ABSTRACT Staphylococcus aureus TF2758 is a clinical isolate from an atheroma and a super-biofilm-elaborating/polysaccharide intercellular adhesin (PIA)/poly-N-acetylglucosamine (PNAG)-overproducing strain (L. Shrestha et al., Microbiol Immunol 60:148–159, 2016, https://doi.org/10.1111/1348-0421.1235...

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Autores principales:	Liansheng Yu, Junzo Hisatsune, Ikue Hayashi, Nobuyuki Tatsukawa, Yusuke Sato’o, Emiri Mizumachi, Fuminori Kato, Hideki Hirakawa, Gerald B. Pier, Motoyuki Sugai
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/97e22f4450f746238c980073c48910e0
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spelling	oai:doaj.org-article:97e22f4450f746238c980073c48910e02021-11-15T15:51:07ZA Novel Repressor of the <italic toggle="yes">ica</italic> Locus Discovered in Clinically Isolated Super-Biofilm-Elaborating <italic toggle="yes">Staphylococcus aureus</italic>10.1128/mBio.02282-162150-7511https://doaj.org/article/97e22f4450f746238c980073c48910e02017-03-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02282-16https://doaj.org/toc/2150-7511ABSTRACT Staphylococcus aureus TF2758 is a clinical isolate from an atheroma and a super-biofilm-elaborating/polysaccharide intercellular adhesin (PIA)/poly-N-acetylglucosamine (PNAG)-overproducing strain (L. Shrestha et al., Microbiol Immunol 60:148–159, 2016, https://doi.org/10.1111/1348-0421.12359 ). A microarray analysis and DNA genome sequencing were performed to identify the mechanism underlying biofilm overproduction by TF2758. We found high transcriptional expression levels of a 7-gene cluster (satf2580 to satf2586) and the ica operon in TF2758. Within the 7-gene cluster, a putative transcriptional regulator gene designated rob had a nonsense mutation that caused the truncation of the protein. The complementation of TF2758 with rob from FK300, an rsbU-repaired derivative of S. aureus strain NCTC8325-4, significantly decreased biofilm elaboration, suggesting a role for rob in this process. The deletion of rob in non-biofilm-producing FK300 significantly increased biofilm elaboration and PIA/PNAG production. In the search for a gene(s) in the 7-gene cluster for biofilm elaboration controlled by rob, we identified open reading frame (ORF) SAOUHSC_2898 (satf2584). Our results suggest that ORF SAOUHSC_2898 (satf2584) and icaADBC are required for enhanced biofilm elaboration and PIA/PNAG production in the rob deletion mutant. Rob bound to a palindromic sequence within its own promoter region. Furthermore, Rob recognized the TATTT motif within the icaR-icaA intergenic region and bound to a 25-bp DNA stretch containing this motif, which is a critically important short sequence regulating biofilm elaboration in S. aureus. Our results strongly suggest that Rob is a long-sought repressor that recognizes and binds to the TATTT motif and is an important regulator of biofilm elaboration through its control of SAOUHSC_2898 (SATF2584) and Ica protein expression in S. aureus. IMPORTANCE During the search for molecular mechanisms underlying biofilm overproduction of Staphylococcus aureus TF2758, we found a putative transcriptional regulator gene designated rob within a 7-gene cluster showing a high transcriptional expression level by microarray analysis. The deletion of rob in non-biofilm-producing FK300, an rsbU-repaired derivative of NCTC8325-4, significantly increased biofilm elaboration and PIA/PNAG production. The search for a gene(s) in the 7-gene cluster for biofilm elaboration controlled by rob identified ORF SAOUHSC_2898. Besides binding to its own promoter region to control ORF SAOUHSC_2898 expression, Rob recognized the TATTT motif within the icaR-icaA intergenic region and bound to a 25-bp DNA stretch containing this motif, which is a critically important short sequence regulating biofilm elaboration in S. aureus. Our results strongly suggest that Rob is a long-sought repressor that recognizes and binds to the TATTT motif and is a new important regulator of biofilm elaboration through its control of SAOUHSC_2898 and Ica protein expression in S. aureus.Liansheng YuJunzo HisatsuneIkue HayashiNobuyuki TatsukawaYusuke Sato’oEmiri MizumachiFuminori KatoHideki HirakawaGerald B. PierMotoyuki SugaiAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 8, Iss 1 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Liansheng Yu Junzo Hisatsune Ikue Hayashi Nobuyuki Tatsukawa Yusuke Sato’o Emiri Mizumachi Fuminori Kato Hideki Hirakawa Gerald B. Pier Motoyuki Sugai A Novel Repressor of the <italic toggle="yes">ica</italic> Locus Discovered in Clinically Isolated Super-Biofilm-Elaborating <italic toggle="yes">Staphylococcus aureus</italic>
description	ABSTRACT Staphylococcus aureus TF2758 is a clinical isolate from an atheroma and a super-biofilm-elaborating/polysaccharide intercellular adhesin (PIA)/poly-N-acetylglucosamine (PNAG)-overproducing strain (L. Shrestha et al., Microbiol Immunol 60:148–159, 2016, https://doi.org/10.1111/1348-0421.12359 ). A microarray analysis and DNA genome sequencing were performed to identify the mechanism underlying biofilm overproduction by TF2758. We found high transcriptional expression levels of a 7-gene cluster (satf2580 to satf2586) and the ica operon in TF2758. Within the 7-gene cluster, a putative transcriptional regulator gene designated rob had a nonsense mutation that caused the truncation of the protein. The complementation of TF2758 with rob from FK300, an rsbU-repaired derivative of S. aureus strain NCTC8325-4, significantly decreased biofilm elaboration, suggesting a role for rob in this process. The deletion of rob in non-biofilm-producing FK300 significantly increased biofilm elaboration and PIA/PNAG production. In the search for a gene(s) in the 7-gene cluster for biofilm elaboration controlled by rob, we identified open reading frame (ORF) SAOUHSC_2898 (satf2584). Our results suggest that ORF SAOUHSC_2898 (satf2584) and icaADBC are required for enhanced biofilm elaboration and PIA/PNAG production in the rob deletion mutant. Rob bound to a palindromic sequence within its own promoter region. Furthermore, Rob recognized the TATTT motif within the icaR-icaA intergenic region and bound to a 25-bp DNA stretch containing this motif, which is a critically important short sequence regulating biofilm elaboration in S. aureus. Our results strongly suggest that Rob is a long-sought repressor that recognizes and binds to the TATTT motif and is an important regulator of biofilm elaboration through its control of SAOUHSC_2898 (SATF2584) and Ica protein expression in S. aureus. IMPORTANCE During the search for molecular mechanisms underlying biofilm overproduction of Staphylococcus aureus TF2758, we found a putative transcriptional regulator gene designated rob within a 7-gene cluster showing a high transcriptional expression level by microarray analysis. The deletion of rob in non-biofilm-producing FK300, an rsbU-repaired derivative of NCTC8325-4, significantly increased biofilm elaboration and PIA/PNAG production. The search for a gene(s) in the 7-gene cluster for biofilm elaboration controlled by rob identified ORF SAOUHSC_2898. Besides binding to its own promoter region to control ORF SAOUHSC_2898 expression, Rob recognized the TATTT motif within the icaR-icaA intergenic region and bound to a 25-bp DNA stretch containing this motif, which is a critically important short sequence regulating biofilm elaboration in S. aureus. Our results strongly suggest that Rob is a long-sought repressor that recognizes and binds to the TATTT motif and is a new important regulator of biofilm elaboration through its control of SAOUHSC_2898 and Ica protein expression in S. aureus.
format	article
author	Liansheng Yu Junzo Hisatsune Ikue Hayashi Nobuyuki Tatsukawa Yusuke Sato’o Emiri Mizumachi Fuminori Kato Hideki Hirakawa Gerald B. Pier Motoyuki Sugai
author_facet	Liansheng Yu Junzo Hisatsune Ikue Hayashi Nobuyuki Tatsukawa Yusuke Sato’o Emiri Mizumachi Fuminori Kato Hideki Hirakawa Gerald B. Pier Motoyuki Sugai
author_sort	Liansheng Yu
title	A Novel Repressor of the <italic toggle="yes">ica</italic> Locus Discovered in Clinically Isolated Super-Biofilm-Elaborating <italic toggle="yes">Staphylococcus aureus</italic>
title_short	A Novel Repressor of the <italic toggle="yes">ica</italic> Locus Discovered in Clinically Isolated Super-Biofilm-Elaborating <italic toggle="yes">Staphylococcus aureus</italic>
title_full	A Novel Repressor of the <italic toggle="yes">ica</italic> Locus Discovered in Clinically Isolated Super-Biofilm-Elaborating <italic toggle="yes">Staphylococcus aureus</italic>
title_fullStr	A Novel Repressor of the <italic toggle="yes">ica</italic> Locus Discovered in Clinically Isolated Super-Biofilm-Elaborating <italic toggle="yes">Staphylococcus aureus</italic>
title_full_unstemmed	A Novel Repressor of the <italic toggle="yes">ica</italic> Locus Discovered in Clinically Isolated Super-Biofilm-Elaborating <italic toggle="yes">Staphylococcus aureus</italic>
title_sort	novel repressor of the <italic toggle="yes">ica</italic> locus discovered in clinically isolated super-biofilm-elaborating <italic toggle="yes">staphylococcus aureus</italic>
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/97e22f4450f746238c980073c48910e0
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A Novel Repressor of the <italic toggle="yes">ica</italic> Locus Discovered in Clinically Isolated Super-Biofilm-Elaborating <italic toggle="yes">Staphylococcus aureus</italic>

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