Multiple factors regulate the expression of sufCDSUB in Streptococcus mutans .

Autor:	Ellepola K; Department of Oral and Craniofacial Biology, School of Dentistry, Louisiana State University Health Sciences Center, New Orleans, LA, United States., Guillot LC; Department of Oral and Craniofacial Biology, School of Dentistry, Louisiana State University Health Sciences Center, New Orleans, LA, United States., Comeaux B; Department of Oral and Craniofacial Biology, School of Dentistry, Louisiana State University Health Sciences Center, New Orleans, LA, United States., Han Y; Department of Biology, Middlebury College, Middlebury, VT, United States., Kajfasz JK; Department of Oral Biology, School of Dentistry, University of Florida, Gainesville, FL, United States., Bitoun JP; Department of Microbiology, Tulane University, New Orleans, LA, United States., Spatafora G; Department of Biology, Middlebury College, Middlebury, VT, United States., Lemos JA; Department of Oral Biology, School of Dentistry, University of Florida, Gainesville, FL, United States., Wen ZT; Department of Oral and Craniofacial Biology, School of Dentistry, Louisiana State University Health Sciences Center, New Orleans, LA, United States.; Department of Microbiology, Immunology and Parasitology, School of Medicine, Louisiana State University Health Sciences Center, New Orleans, LA, United States.
Jazyk:	angličtina
Zdroj:	Frontiers in cellular and infection microbiology [Front Cell Infect Microbiol] 2024 Nov 27; Vol. 14, pp. 1499476. Date of Electronic Publication: 2024 Nov 27 (Print Publication: 2024).
DOI:	10.3389/fcimb.2024.1499476
Abstrakt:	Introduction: The sufCDSUB gene cluster, encoding the sole iron-sulfur (Fe-S) cluster assembly system in S. mutans , was recently shown to be up-regulated in response to oxidative stressors and Fe limitation. Methods: In this study, luciferase reporter fusion assays, electrophoretic gel mobility shift assays (EMSA) and in vitro transcription assays (IVT) were used to dissect the cis- and trans- acting factors that regulate the expression of sufCDSUB . Results and Discussion: Results showed deletion of perR , for the only Fur-family transcriptional regulator in S. mutans , resulted in >5-fold increases in luciferase activity under the control of the sufCDSUB promoter (P<0.01), as compared to the parent strain, UA159 when the reporter strains were grown in medium with no supplemental iron. Site-directed mutagenesis of a PerR-box in the promoter region led to elevation of the reporter activity by >1.6-fold ( P <0.01). In an EMSA, recombinant PerR (rPerR) was shown to bind to the cognate sufCDSUB promoter leading to mobility retardation. On the other hand, the reporter activity was increased by >84-fold (P<0.001) in response to the addition of cysteine at 4 mM to the culture medium. Deletion of cysR , for a LysR-type of transcriptional regulator, led to reduction of the reporter activity by >11.6-fold ( P <0.001). Addition of recombinant CysR (rCysR) to an EMSA caused mobility shift of the sufCDSUB promoter probe, indicative of rCysR-promoter interaction, and rCysR was shown to enhance sufC transcription under the direction of sufCDSUB promoter in vitro . These results suggest that multiple factors are involved in the regulation of sufCDSUB expression in response to environmental cues, including cysteine and Fe availability, consistent with the important role of sufCDSUB in S. mutans physiology. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision. (Copyright © 2024 Ellepola, Guillot, Comeaux, Han, Kajfasz, Bitoun, Spatafora, Lemos and Wen.)
Databáze:	MEDLINE
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