Analysis of Haloferax mediterranei Lrp Transcriptional Regulator
| Autor: | Laura Matarredona, Mónica Camacho, Maria-Jose Garcia-Bonete, María José Bonete, Basilio Zafrilla, Julia Esclapez, Belén Esquerra |
|---|---|
| Přispěvatelé: | Universidad de Alicante. Departamento de Agroquímica y Bioquímica, Universidad de Alicante. Departamento de Fisiología, Genética y Microbiología, Biotecnología de Extremófilos (BIOTECEXTREM), Microbiología Molecular |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Lrp 030106 microbiology Haloferax mediterranei his-tag homologous overexpression Western blot QH426-470 Stress Microbiología 03 medical and health sciences stress Transcriptional regulation Genetics Gene Genetics (clinical) Regulation of gene expression Reporter gene Homologous overexpression western blot biology Chemistry Promoter Bioquímica y Biología Molecular biology.organism_classification Archaea 030104 developmental biology Biochemistry β-galactosidase assay Haloarchaea His-tag |
| Zdroj: | Genes Volume 12 Issue 6 Genes, Vol 12, Iss 802, p 802 (2021) RUA. Repositorio Institucional de la Universidad de Alicante Universidad de Alicante (UA) |
| ISSN: | 2073-4425 |
| DOI: | 10.3390/genes12060802 |
| Popis: | Haloferax mediterranei is an extremely halophilic archaeon, able to live in hypersaline environments with versatile nutritional requirements, whose study represents an excellent basis in the field of biotechnology. The transcriptional machinery in Archaea combines the eukaryotic basal apparatus and the bacterial regulation mechanisms. However, little is known about molecular mechanisms of gene expression regulation compared with Bacteria, particularly in Haloarchaea. The genome of Hfx. mediterranei contains a gene, lrp (HFX_RS01210), which encodes a transcriptional factor belonging to Lrp/AsnC family. It is located downstream of the glutamine synthetase gene (HFX_RS01205), an enzyme involved in ammonium assimilation and amino acid metabolism. To study this transcriptional factor more deeply, the lrp gene has been homologously overexpressed and purified under native conditions by two chromatographic steps, namely nickel affinity and gel filtration chromatography, showing that Lrp behaves asa tetrameric protein of approximately 67 kDa. Its promoter region has been characterized under different growth conditions using bgaH as a reporter gene. The amount of Lrp protein was also analyzed by Western blotting in different nitrogen sources and under various stress conditions. To sum up, regarding its involvement in the nitrogen cycle, it has been shown that its expression profile does not change in response to the nitrogen sources tested. Differences in its expression pattern have been observed under different stress conditions, such as in the presence of hydrogen peroxide or heavy metals. According to these results, the Lrp seems to be involved in a general response against stress factors, acting as a first-line transcriptional regulator. This research was funded by Universidad de Alicante, VIGROB-016. |
| Databáze: | OpenAIRE |
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