Fructose-1-kinase has pleiotropic roles in Escherichia coli .
Chamitha WeeramangeCindy MenjivarPierce T O'NeilSamir El QaidiKelly S HarrisonSarah MeinhardtCole L BirdShwetha SreenivasanPhilip R HardwidgeAron W FentonP Scott HeftyJeffrey L BoseLiskin Swint-KrusePublished in: bioRxiv : the preprint server for biology (2023)
In Escherichia coli , the master transcription regulator Catabolite Repressor Activator (Cra) regulates >100 genes in central metabolism. Cra binding to DNA is allosterically regulated by binding to fructose-1-phosphate (F-1-P), but the only documented source of F-1-P is from the concurrent import and phosphorylation of exogenous fructose. Thus, many have proposed that fructose-1,6-bisphosphate (F-1,6-BP) is also a physiological regulatory ligand. However, the role of F-1,6-BP has been widely debated. Here, we report that the E. coli enzyme fructose-1-kinase (FruK) can carry out its "reverse" reaction under physiological substrate concentrations to generate F-1-P from F-1,6-BP. We further show that FruK directly binds Cra with nanomolar affinity and forms higher order, heterocomplexes. Growth assays with a Δ fruK strain and fruK complementation show that FruK has a broader role in metabolism than fructose catabolism. The Δ fruK strain also alters biofilm formation. Since fruK itself is repressed by Cra, these newly-reported events add layers to the dynamic regulation of E. coli central metabolism that occur in response to changing nutrients. These findings might have wide-spread relevance to other γ-proteobacteria, which conserve both Cra and FruK.
Keyphrases
- escherichia coli
- biofilm formation
- pseudomonas aeruginosa
- staphylococcus aureus
- protein kinase
- klebsiella pneumoniae
- high throughput
- risk assessment
- tyrosine kinase
- gene expression
- immune response
- single molecule
- cystic fibrosis
- heavy metals
- single cell
- circulating tumor
- dna methylation
- inflammatory response
- circulating tumor cells
- locally advanced
- capillary electrophoresis