2026-06-14T03:33:51Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/160962025-05-08T13:09:44Zcom_20.500.12105_2052com_20.500.12105_2051col_20.500.12105_19609col_20.500.12105_25179
Repisalud
author
de Vasconcelos Júnior, Antônio Alexandre
author
Tirado-Velez, JM
author
Martin-Galiano, Antonio Javier
author
Megías, Diego
author
Ferrandiz-Avellano, Maria-Jose
author
Hernández, Pablo
author
Amblar, Monica
author
de la Campa, Adela G
funder
Agencia Estatal de Investigación (España)
funder
Ministerio de Economía e Innovación (España)
funder
Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF)
2023-05-18T11:36:40Z
2023-05-18T11:36:40Z
2023-03-22
Int J Mol Sci. 2023 Mar 22;24(6):5973.
10.3390/ijms24065973
1422-0067
International journal of molecular sciences
36983048
http://hdl.handle.net/20.500.12105/16096
The DNA topoisomerases gyrase and topoisomerase I as well as the nucleoid-associated protein HU maintain supercoiling levels in Streptococcus pneumoniae, a main human pathogen. Here, we characterized, for the first time, a topoisomerase I regulator protein (StaR). In the presence of sub-inhibitory novobiocin concentrations, which inhibit gyrase activity, higher doubling times were observed in a strain lacking staR, and in two strains in which StaR was over-expressed either under the control of the ZnSO4-inducible PZn promoter (strain ΔstaRPZnstaR) or of the maltose-inducible PMal promoter (strain ΔstaRpLS1ROMstaR). These results suggest that StaR has a direct role in novobiocin susceptibility and that the StaR level needs to be maintained within a narrow range. Treatment of ΔstaRPZnstaR with inhibitory novobiocin concentrations resulted in a change of the negative DNA supercoiling density (σ) in vivo, which was higher in the absence of StaR (σ = -0.049) than when StaR was overproduced (σ = -0.045). We have located this protein in the nucleoid by using super-resolution confocal microscopy. Through in vitro activity assays, we demonstrated that StaR stimulates TopoI relaxation activity, while it has no effect on gyrase activity. Interaction between TopoI and StaR was detected both in vitro and in vivo by co-immunoprecipitation. No alteration of the transcriptome was associated with StaR amount variation. The results suggest that StaR is a new streptococcal nucleoid-associated protein that activates topoisomerase I activity by direct protein-protein interaction.
eng
DNA gyrase
DNA supercoiling
DNA topoisomerase I
StaR
Nucleoid-associated proteins
Supercoiling homeostasis
Supercoiling regulation
Topsoisomerase regulator
StaR Is a Positive Regulator of Topoisomerase I Activity Involved in Supercoiling Maintenance in Streptococcus pneumoniae
research article
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StaR_IsPositiveRegulatorTopoisomeraseI_2023.pdf
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StaR_IsPositiveRegulatorTopoisomeraseI_2023.pdf.txt