The lumina of the endoplasmic reticulum and Golgi apparatus are the subcellular sites where glycosylation, sulfation, and phosphorylation of secretory and membrane-bound proteins, proteoglycans, and lipids occur. Nucleotide sugars, nucleotide sulfate, and ATP are substrates in the above reactions and must first be translocated from the cytosol into the lumen of these organelles. Translocation of these nucleotide derivatives is mediated by highly specific transporters, which are antiporters with the corresponding nucleoside monophosphate, as shown by genetic and biochemical approaches in mammals and yeast. Studies with mammalian, yeast, and protozoa mutants have shown that a defect in a specific translocator results in selective impairments of glycosylation of proteins, lipids and proteoglycans in vivo. Several of these transporters have been purified, cloned, and found to encode very hydrophobic proteins with multitransmembrane domains. Experiments with yeast and mammalian cells demonstrate that these transporters play a regulatory role in posttranslational modifications.