Hexose phosphorylation is an essential step of sugar metabolism. Only two classes of glucose and fructose phosphorylating enzymes, hexokinases (HXK) and fructokinases (FRK), have been found in plants. Tomato (Lycopersicon esculentum Mill.) is the only plant species from which four HXK and four FRK genes have been identified and characterised. One HXK and one FRK isozyme are located within plastids. The other three HXK isozymes are associated with the mitochondria, and the other three FRK isozymes are dispersed in the cytosol. These differences in location suggest that the cytoplasmic HXK and FRK have distinct roles to play in sugar metabolism. The specific roles of each of the HXK and FRK genes have been investigated using transgenic plants with modified expression of the genes. Sugar signalling effects were obtained with modified expression of the mitochondria associated HXK. In contrast, modified expression of the cytosolic FRK affected fructose metabolism rather than sugar signalling. Future research efforts will aim to determining the roles of specific hexose phosphorylating enzymes in tomato plants, the source of the hexose monomers to be phosphorylated, and their intracellular trafficking route.