Influence of iron and heme availability on level of mRNA encoding P. endodontalis HmuY protein (RT-qPCR data)

The upload contains raw data files for the article "Porphyromonas endodontalis HmuY differentially participates in heme acquisition compared to the Porphyromonas gingivalis and Tannerella forsythia hemophore-like proteins" which shows the influence of iron and heme availability on expression of Porphyromonas endodontalis hmuY gene (hmuYPe) using RT-qPCR. These data are not present in the supplementary data of the publication.

P. endodontalis ATCC 35406 (Argenta, Poznań, Poland) was grown on Schaedler blood agar (ABA) plates (Argenta) at 37°C for 6 days under anaerobic conditions (80% N2, 10% H2 and 10% CO2; Whitley A35 anaerobic workstation; Bingley, UK). Then, bacteria were used to inoculate a liquid basal medium (BM) prepared of 3% trypticase soy broth (Becton Dickinson, Sparks, MD, USA) and 0.5% yeast extract (Biomaxima, Lublin, Poland), 0.05% L-cysteine (Carl Roth, Karlsruhe, Germany), 0.5 mg/l menadione (Sigma-Aldrich, St. Louis, MO, USA) (BM medium). To grow bacteria under optimal conditions, 7.7 μM hemin chloride (Fluka, Munich, Germany) was added to form iron and heme rich conditions (Hm medium). To starve bacteria of iron and heme, heme was not added and the medium was supplemented with 160 μM 2,2-dipyridyl (Sigma-Aldrich) to chelate free iron (DIP medium). The optical density at 600 nm (OD600) at the beginning of the liquid culture was at least 0.5. Determination of hmuYPe gene expression was examined at the transcript level by RT-qPCR using 16S rRNA as a reference gene. mRNA levels were determined in bacteria starved of iron and heme compared to bacteria grown in rich iron and heme conditions. Bacteria from 1 ml of culture were centrifuged and used for RNA isolation as previously described (Śmiga and Olczak, 2024). qPCR was performed using SensiFAST SYBR no-ROX Kit (Bioline, London, UK) and LightCycler 96 (Roche, Basel, Switzerland). The Word document (Data description.docx) contains a description of files.

The Word document (Data description.docx) contains a description of files.