title: Characterization of a novel two-component regulatory system, HptRS, the regulator for the hexose phosphate transport system in Staphylococcus aureus.
abstract: Hexose phosphate is an important carbon source within the cytoplasm of host cells. Bacterial pathogens that invade, survive, and multiply within various host epithelial cells exploit hexose phosphates from the host cytoplasm through the hexose phosphate transport (HPT) system to gain energy and synthesize cellular components. In Escherichia coli, the HPT system consists of a two-component regulatory system (UhpAB) and a phosphate sensor protein (UhpC) that tightly regulate expression of a hexose phosphate transporter (UhpT). Although growing evidence suggests that Staphylococcus aureus also can invade, survive, and multiply within various host epithelial cells, the genetic elements involved in the HPT system in S. aureus have not been characterized yet. In this study, we identified and characterized the HPT system in S. aureus that includes the hptRS (a novel two-component regulatory system), the hptA (a putative phosphate sensor), and the uhpT (a hexose phosphate transporter) genes. The hptA, hptRS, and uhpT markerless deletion mutants were generated by an allelic replacement method using a modified pMAD-CM-GFPuv vector system. We demonstrated that both hptA and hptRS are required to positively regulate transcription of uhpT in response to extracellular phosphates, such as glycerol-3-phosphate (G3P), glucose-6-phosphate (G6P), and fosfomycin. Mutational studies revealed that disruption of the hptA, hptRS, or uhpT gene impaired the growth of bacteria when the available carbon source was limited to G6P, impaired survival/multiplication within various types of host cells, and increased resistance to fosfomycin. The results of this study suggest that the HPT system plays an important role in adaptation of S. aureus within the host cells and could be an important target for developing novel antistaphylococcal therapies.
authors: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA.
authors: Joo Youn
authors: JY
authors: Park
authors: Animal Diseases Diagnostic Division, Quarantine and Inspection Agency, Anyang, Gyeonggi, Republic of Korea.
authors: Jong Wan
authors: JW
authors: Kim
authors: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA Department of Microbiology, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
authors: Bo Youn
authors: BY
authors: Moon
authors: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA.
authors: Juyeun
authors: J
authors: Lee
authors: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA.
authors: Ye Ji
authors: YJ
authors: Fortin
authors: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA.
authors: Frank W
authors: FW
authors: Austin
authors: Division of Infectious Diseases, Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center, Torrance, California, USA The David Geffen School of Medicine at UCLA, Los Angeles, California, USA.
authors: Soo-Jin
authors: SJ
authors: Yang
authors: Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, USA seo@cvm.msstate.edu.
authors: Keun Seok
authors: http://orcid.org/0000-0001-9280-9680
authors: KS
authors: Seo
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics; D001692:Biological Transport / Q000235:genetics; D002460:Cell Line; D024881:Drug Resistance, Bacterial; D004847:Epithelial Cells / Q000382:microbiology; D005578:Fosfomycin / Q000494:pharmacology*; D017353:Gene Deletion; D019298:Glucose-6-Phosphate / Q000378:metabolism; D006601:Hexoses / Q000378:metabolism*; D006801:Humans; D051379:Mice; D009004:Monosaccharide Transport Proteins / Q000235:genetics*; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D006601:Hexoses; D009004:Monosaccharide Transport Proteins; D005578:Fosfomycin; D019298:Glucose-6-Phosphate
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