A tale of two functions: enzymatic activity and translational repression by carboxyltransferase.
Acetyl-CoA Carboxylase catalyzes the first committed step in fatty acid synthesis. Escherichia coli acetyl-CoA carboxylase is composed of biotin carboxylase, carboxyltransferase and biotin carboxyl carrier protein functions. The accA and accD genes that code for the alpha- and beta-subunits, respectively, are not in an operon, yet yield an alpha(2)beta(2) carboxyltransferase. Here, we report that carboxyltransferase regulates its own translation by binding the mRNA encoding its subunits. This interaction is mediated by a zinc finger on the beta-subunit; mutation of the four cysteines to alanine diminished nucleic acid binding and catalytic activity. Carboxyltransferase binds the coding regions of both subunit mRNAs and inhibits translation, an inhibition that is relieved by the substrate acetyl-CoA. mRNA binding reciprocally inhibits catalytic activity. Preferential binding of carboxyltransferase to RNA in situ was shown using fluorescence resonance energy transfer. We propose an unusual regulatory mechanism by which carboxyltransferase acts as a 'dimmer switch' to regulate protein production and catalytic activity, while sensing the metabolic state of the cell through acetyl-CoA concentration.
Meades
Glen
G
Division of Biochemistry and Molecular Biology, Louisiana State University, Baton Rouge, LA 70803, USA.
Benson
Brian K
BK
Grove
Anne
A
Waldrop
Grover L
GL
D000103:Acetyl-CoA Carboxylase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D019878:Carboxyl and Carbamoyl Transferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002384:Catalysis; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D014176:Protein Biosynthesis; D021122:Protein Subunits / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D016335:Zinc Fingers
D029968:Escherichia coli Proteins; D021122:Protein Subunits; D012333:RNA, Messenger; D004247:DNA; D019878:Carboxyl and Carbamoyl Transferases; D000103:Acetyl-CoA Carboxylase