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Johnsen, Ulrike; Ortjohann, Marius; Reinhardt, Andreas; Turner, Jonathan M.; Stratton, Caleb; Weber, Katherine R.; Sanchez, Karol M.; Maupin‐Furlow, Julie; Davies, Christopher; Schönheit, Peter
Molecular microbiology, August 2023, Volume: 120, Issue: 2Journal Article
The haloarchaeon Haloferax volcanii degrades D‐glucose via the semiphosphorylative Entner‐Doudoroff pathway and D‐fructose via a modified Embden‐Meyerhof pathway. Here, we report the identification of GfcR, a novel type of transcriptional regulator that functions as an activator of both D‐glucose and D‐fructose catabolism. We find that in the presence of D‐glucose, GfcR activates gluconate dehydratase, glyceraldehyde‐3‐phosphate dehydrogenase and pyruvate kinase and also acts as activator of the phosphotransferase system and of fructose‐1,6‐bisphosphate aldolase, which are involved in uptake and degradation of D‐fructose. In addition, glyceraldehyde‐3‐phosphate dehydrogenase and pyruvate kinase are activated by GfcR in the presence of D‐fructose and also during growth on D‐galactose and glycerol. Electrophoretic mobility shift assays indicate that GfcR binds directly to promoters of regulated genes. Specific intermediates of the degradation pathways of the three hexoses and of glycerol were identified as inducer molecules of GfcR. GfcR is composed of a phosphoribosyltransferase (PRT) domain with an N‐terminal helix‐turn‐helix motif and thus shows homology to PurR of Gram‐positive bacteria that is involved in the transcriptional regulation of nucleotide biosynthesis. We propose that GfcR of H. volcanii evolved from a PRT‐like enzyme to attain a function as a transcriptional regulator of central sugar catabolic pathways in archaea. A novel transcriptional regulator, GfcR, has been identified that activates the degradation pathways of glucose, fructose, galactose and glycerol in the archaeon Haloferax volcanii. GfcR comprises a phosphoribosyltransferase domain with an N‐terminal helix‐turn‐helix DNA‐binding motif. Activation proceeds via a feedforward mechanism, where specific intermediates of the degradation pathways bind to GfcR and induce transcription of enzymes within those pathways. These data expand our knowledge of transcriptional regulation of central metabolic pathways in archaea.
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