Staphylococcus aureus SarA is a regulatory protein responsive to redox and pH that can support bacteriophage lambda integrase-mediated excision/recombination.
Staphylococcus aureus produces a wide array of virulence factors and causes a correspondingly diverse array of infections. Production of these virulence factors is under the control of a complex network of global regulatory elements, one of which is sarA. sarA encodes a DNA binding protein that is considered to function as a transcription factor capable of acting as either a repressor or an activator. Using competitive ELISA assays, we demonstrate that SarA is present at approximately 50 000 copies per cell, which is not characteristic of classical transcription factors. We also demonstrate that SarA is present at all stages of growth in vitro and is capable of binding DNA with high affinity but that its binding affinity and pattern of shifted complexes in electrophoretic mobility shift assays is responsive to the redox state. We also show that SarA binds to the bacteriophage lambda (lambda) attachment site, attL, producing SarA-DNA complexes similar to intasomes, which consist of bacteriophage lambda integrase, Escherichia coli integration host factor and attL DNA. In addition, SarA stimulates intramolecular excision recombination in the absence of lambda excisionase, a DNA binding accessory protein. Taken together, these data suggest that SarA may function as an architectural accessory protein.
Fujimoto
David F
DF
Department of Biology, San Diego State University, San Diego, CA 92182, USA.
Higginbotham
Robin H
RH
Sterba
Kristen M
KM
Maleki
Soheila J
SJ
Segall
Anca M
AM
Smeltzer
Mark S
MS
Hurlburt
Barry K
BK
D000595:Amino Acid Sequence; D001287:Attachment Sites, Microbiological; D001426:Bacterial Proteins / Q000378:metabolism*; D010582:Bacteriophage lambda / Q000201:enzymology*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000235:genetics / Q000821:virology; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D006863:Hydrogen-Ion Concentration; D019426:Integrases / Q000096:biosynthesis*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011995:Recombination, Genetic; D016415:Sequence Alignment; D013211:Staphylococcus aureus / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C492107:SarA protein, Staphylococcus aureus; D014157:Transcription Factors; D019426:Integrases