Organizational requirements of the saer binding sites for a functional P1 promoter of the sae operon in Staphylococcus aureus

Hoonsik Cho, Do Won Jeong, Chunling Li, Taeok Bae

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

In Staphylococcus aureus, the SaeRS two-component system controls the expression of multiple virulence factors. Of the two promoters in the sae operon, P1 is autoinduced and has two binding sites for the response regulator SaeR. In this study, we examined the organizational requirements of the SaeR binding sites in P1 for transcription activation. Mutational studies showed that both binding sites are essential for binding to phosphorylated SaeR (P-SaeR) and transcription activation. When the 21-bp distance between the centers of the two SaeR binding sites was altered to 26 bp, 31 bp, 36 bp, or 41 bp, only the 31-bp mutant retained approximately 40% of the original promoter activity. When the 21-bp spacing (i.e.,1-bp overlap) between the primary SaeR binding site and the-35 promoter region was altered, all mutant P1 promoters failed to initiate transcription; however, when the first nucleotide of the-35 region was changed from A to T, the mutants with 0-bp or 22-bp spacing showed detectable promoter activity. Although P-SaeR was essential for the binding of RNA polymerase to P1, it was not essential for the binding of the enzyme to the alpha-hemolysin promoter. When the nonoptimal spacing between promoter elements in P1 or the coagulase promoter was altered to the optimal spacing of 17 bp, both promoters failed to initiate transcription. These results suggest that SaeR binding sites are under rather strict organizational restrictions and provide clues for understanding the molecular mechanism of sae-mediated transcription activation.

Original languageEnglish
Pages (from-to)2865-2876
Number of pages12
JournalJournal of Bacteriology
Volume194
Issue number11
DOIs
StatePublished - Jun 2012

Fingerprint

Operon
Staphylococcus aureus
Binding Sites
Transcriptional Activation
Hemolysin Proteins
Coagulase
Virulence Factors
DNA-Directed RNA Polymerases
Genetic Promoter Regions
Nucleotides
Enzymes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Organizational requirements of the saer binding sites for a functional P1 promoter of the sae operon in Staphylococcus aureus. / Cho, Hoonsik; Jeong, Do Won; Li, Chunling; Bae, Taeok.

In: Journal of Bacteriology, Vol. 194, No. 11, 06.2012, p. 2865-2876.

Research output: Contribution to journalArticle

@article{1da3100bed5045478237a122c1d71c48,
title = "Organizational requirements of the saer binding sites for a functional P1 promoter of the sae operon in Staphylococcus aureus",
abstract = "In Staphylococcus aureus, the SaeRS two-component system controls the expression of multiple virulence factors. Of the two promoters in the sae operon, P1 is autoinduced and has two binding sites for the response regulator SaeR. In this study, we examined the organizational requirements of the SaeR binding sites in P1 for transcription activation. Mutational studies showed that both binding sites are essential for binding to phosphorylated SaeR (P-SaeR) and transcription activation. When the 21-bp distance between the centers of the two SaeR binding sites was altered to 26 bp, 31 bp, 36 bp, or 41 bp, only the 31-bp mutant retained approximately 40{\%} of the original promoter activity. When the 21-bp spacing (i.e.,1-bp overlap) between the primary SaeR binding site and the-35 promoter region was altered, all mutant P1 promoters failed to initiate transcription; however, when the first nucleotide of the-35 region was changed from A to T, the mutants with 0-bp or 22-bp spacing showed detectable promoter activity. Although P-SaeR was essential for the binding of RNA polymerase to P1, it was not essential for the binding of the enzyme to the alpha-hemolysin promoter. When the nonoptimal spacing between promoter elements in P1 or the coagulase promoter was altered to the optimal spacing of 17 bp, both promoters failed to initiate transcription. These results suggest that SaeR binding sites are under rather strict organizational restrictions and provide clues for understanding the molecular mechanism of sae-mediated transcription activation.",
author = "Hoonsik Cho and Jeong, {Do Won} and Chunling Li and Taeok Bae",
year = "2012",
month = "6",
doi = "10.1128/JB.06771-11",
language = "English",
volume = "194",
pages = "2865--2876",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "11",

}

TY - JOUR

T1 - Organizational requirements of the saer binding sites for a functional P1 promoter of the sae operon in Staphylococcus aureus

AU - Cho, Hoonsik

AU - Jeong, Do Won

AU - Li, Chunling

AU - Bae, Taeok

PY - 2012/6

Y1 - 2012/6

N2 - In Staphylococcus aureus, the SaeRS two-component system controls the expression of multiple virulence factors. Of the two promoters in the sae operon, P1 is autoinduced and has two binding sites for the response regulator SaeR. In this study, we examined the organizational requirements of the SaeR binding sites in P1 for transcription activation. Mutational studies showed that both binding sites are essential for binding to phosphorylated SaeR (P-SaeR) and transcription activation. When the 21-bp distance between the centers of the two SaeR binding sites was altered to 26 bp, 31 bp, 36 bp, or 41 bp, only the 31-bp mutant retained approximately 40% of the original promoter activity. When the 21-bp spacing (i.e.,1-bp overlap) between the primary SaeR binding site and the-35 promoter region was altered, all mutant P1 promoters failed to initiate transcription; however, when the first nucleotide of the-35 region was changed from A to T, the mutants with 0-bp or 22-bp spacing showed detectable promoter activity. Although P-SaeR was essential for the binding of RNA polymerase to P1, it was not essential for the binding of the enzyme to the alpha-hemolysin promoter. When the nonoptimal spacing between promoter elements in P1 or the coagulase promoter was altered to the optimal spacing of 17 bp, both promoters failed to initiate transcription. These results suggest that SaeR binding sites are under rather strict organizational restrictions and provide clues for understanding the molecular mechanism of sae-mediated transcription activation.

AB - In Staphylococcus aureus, the SaeRS two-component system controls the expression of multiple virulence factors. Of the two promoters in the sae operon, P1 is autoinduced and has two binding sites for the response regulator SaeR. In this study, we examined the organizational requirements of the SaeR binding sites in P1 for transcription activation. Mutational studies showed that both binding sites are essential for binding to phosphorylated SaeR (P-SaeR) and transcription activation. When the 21-bp distance between the centers of the two SaeR binding sites was altered to 26 bp, 31 bp, 36 bp, or 41 bp, only the 31-bp mutant retained approximately 40% of the original promoter activity. When the 21-bp spacing (i.e.,1-bp overlap) between the primary SaeR binding site and the-35 promoter region was altered, all mutant P1 promoters failed to initiate transcription; however, when the first nucleotide of the-35 region was changed from A to T, the mutants with 0-bp or 22-bp spacing showed detectable promoter activity. Although P-SaeR was essential for the binding of RNA polymerase to P1, it was not essential for the binding of the enzyme to the alpha-hemolysin promoter. When the nonoptimal spacing between promoter elements in P1 or the coagulase promoter was altered to the optimal spacing of 17 bp, both promoters failed to initiate transcription. These results suggest that SaeR binding sites are under rather strict organizational restrictions and provide clues for understanding the molecular mechanism of sae-mediated transcription activation.

UR - http://www.scopus.com/inward/record.url?scp=84864004481&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84864004481&partnerID=8YFLogxK

U2 - 10.1128/JB.06771-11

DO - 10.1128/JB.06771-11

M3 - Article

VL - 194

SP - 2865

EP - 2876

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 11

ER -