The review examines the mechanisms regulating the activities of the two key enzymes determining rates of glucose and fatty acid oxidation, i.e., the pyruvate dehydrogenase (PDH) complex and the carnitine palmitoyltransferase (CPT) system. The review also evaluates the regulatory importance of gene expression in the control of tissue fuel selection within the context of substrate competition between glucose and fatty acids. It identifies a strong indirect input of nutrient‐gene interactions in the control of pyruvate oxidation through the regulated provision of pyruvate as a substrate for PDH and as an inhibitor of PDH kinase. Nutrient‐gene interactions are also identified in relation to the regulation of CPT I activity by malonyl‐CoA (inhibitor) and by the provision of long‐chain acyl‐CoA (substrate/activator), the latter via the hydrolysis of plasma or tissue triacylglycerol (by lipoprotein lipase and hormone‐sensitive lipase, respectively). We discuss how such regulation is reinforced by long‐term modulation of PDH kinase‐specific activity and CPT I maximal activity. We also explore the role of mechanisms operating at the levels of the PDH complex and the CPT system that act to promote and accelerate a switch in fuel utilization once a committed change in nutrient supply has been established. In particular, we discuss the regulatory influences exerted by altered sensitivities of PDH kinase to inhibition by pyruvate and CPT I to inhibition by malonyl‐CoA, respectively.— Sugden, M. C., Holness, M. J. Interactive regulation of the pyruvate dehydrogenase complex and the carnitine palmitoyltransferase system.FASEB J.8: 54‐61; 1994.