Photorhabdus lux- operon heat shock-like regulation

V V Fomin; S V Bazhenov; O V Kononchuk; V O Matveeva; A P Zarubina; S E Spiridonov; I V Manukhov

doi:10.1016/j.heliyon.2023.e14527

Photorhabdus lux- operon heat shock-like regulation

Heliyon. 2023 Mar 11;9(3):e14527. doi: 10.1016/j.heliyon.2023.e14527. eCollection 2023 Mar.

Authors

V V Fomin^{1

2}, S V Bazhenov¹, O V Kononchuk^{1

2}, V O Matveeva¹, A P Zarubina³, S E Spiridonov⁴, I V Manukhov¹

Affiliations

¹ Laboratory of Molecular Genetics, Moscow Institute of Physics and Technology, Institutsky Lane 9, Dolgoprudny, Moscow Region, 141700, Russian Federation.
² Laboratory of Microbiology, BIOTECH University, Volokolamskoe Highway 11, Moscow 125080, Russian Federation.
³ Biological Faculty, Lomonosov Moscow State University, Vorob'evy Gory, Moscow, 119992, Russian Federation.
⁴ Centre of Parasitology, A.N. Severtsov Institute of Ecology and Evolution, Russian Academy of Sciences, Leninskii Prospect, 33, Moscow, 119071, Russian Federation.

Abstract

For decades, transcription of Photorhabdus luminescens lux-operon was considered being constitutive. Therefore, this lux-operon has been used for measurements in non-specific bacterial luminescent biosensors. Here, the expression of Photorhabdus lux-operon under high temperature was studied. The expression was researched in the natural strain Photorhabdus temperata and in the heterologous system of Escherichia coli. P. temperata FV2201 bacterium was isolated from soil in the Moscow region (growth optimum 28 °C). We showed that its luminescence significantly increases when the temperature rises to 34 °C. The increase in luminescence is associated with an increase in the transcription of luxCDABE genes, which was confirmed by RT-PCR. The promoter of the lux-operon of the related bacterium P. luminescens ZM1 from the forests of Moldova, being cloned in the heterologous system of E. coli, is activated when the temperature rises from room temperature to 42 °C. When heat shock is caused by ethanol addition, transcription of lux-operon increases only in the natural strain of P. temperata, but not in the heterologous system of E. coli cells. In addition, the activation of the lux-operon of P. luminescens persists in E. coli strains deficient in both the rpoH and rpoE genes. These results indicate the presence of sigma 32 and sigma 24 independent heat-shock-like mechanism of regulation of the lux-operon of P. luminescens in the heterologous E. coli system.

Keywords: Heat shock; Lux-operon; P. temperata; Promoter.