title: Novel activities of glycolytic enzymes in Bacillus subtilis: interactions with essential proteins involved in mRNA processing.
abstract: Glycolysis is one of the most important metabolic pathways in heterotrophic organisms. Several genes encoding glycolytic enzymes are essential in many bacteria even under conditions when neither glycolytic nor gluconeogenic activities are required. In this study, a screening for in vivo interaction partners of glycolytic enzymes of the soil bacterium Bacillus subtilis was used to provide a rationale for essentiality of glycolytic enzymes. Glycolytic enzymes proved to be in close contact with several other proteins, among them a high proportion of essential proteins. Among these essential interaction partners, other glycolytic enzymes were most prominent. Two-hybrid studies confirmed interactions of phosphofructokinase with phosphoglyceromutase and enolase. Such a complex of glycolytic enzymes might allow direct substrate channeling of glycolytic intermediates. Moreover we found associations of glycolytic enzymes with several proteins known or suspected to be involved in RNA processing and degradation. One of these proteins, Rny (YmdA), which has so far not been functionally characterized, is required for the processing of the mRNA of the glycolytic gapA operon. Two-hybrid analyses confirmed the interactions between the glycolytic enzymes phosphofructokinase and enolase and the enzymes involved in RNA processing, RNase J1, Rny, and polynucleotide phosphorylase. Moreover RNase J1 interacts with its homologue RNase J2. We suggest that this complex of mRNA processing and glycolytic enzymes is the B. subtilis equivalent of the RNA degradosome. Our findings suggest that the functional interaction of glycolytic enzymes with essential proteins may be the reason why they are indispensable.
authors.affiliation: Department of General Microbiology, Georg-August-University Göttingen, Grisebachstrasse 8, D-37077 Göttingen, Germany.
authors.forename: Fabian M
authors.initials: FM
authors.lastname: Commichau
authors.forename: Fabian M
authors.initials: FM
authors.lastname: Rothe
authors.forename: Christina
authors.initials: C
authors.lastname: Herzberg
authors.forename: Eva
authors.initials: E
authors.lastname: Wagner
authors.forename: Daniel
authors.initials: D
authors.lastname: Hellwig
authors.forename: Martin
authors.initials: M
authors.lastname: Lehnik-Habrink
authors.forename: Elke
authors.initials: E
authors.lastname: Hammer
authors.forename: Uwe
authors.initials: U
authors.lastname: Völker
authors.forename: Jörg
authors.initials: J
authors.lastname: Stülke
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology*; D001483:Base Sequence; D015152:Blotting, Northern; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D006019:Glycolysis; D025221:Phosphofructokinases / Q000378:metabolism*; D010736:Phosphoglycerate Mutase / Q000378:metabolism*; D010751:Phosphopyruvate Hydratase / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger; D025221:Phosphofructokinases; D010751:Phosphopyruvate Hydratase; D010736:Phosphoglycerate Mutase