The beta-1,4-galactosyltransferase (GT; EC 2.4.1.90) is localized in the trans-cisternae of the Golgi apparatus where it catalyzes the transfer of galactose from UDP-galactose to the N-acetylglucosamine residue of secretory and membrane-bound glycoproteins. Given the potential role of GT in cell-cell interaction and the fact that numerous cell surface events occur during cell growth we studied the possible relationship between GT expression and 3T3 cell growth. The level of GT mRNA increases 3--4-fold 2 h after serum-stimulation of quiescent 3T3 cells. Protein biosynthesis inhibitors like cycloheximide and anisomycin superinduce GT mRNA expression. Concomitant with this increase is an observed rise in the level of GT protein as well as an increase in overall GT enzymatic activity. Antibody-binding studies and direct enzyme assays of intact cells, along with subcellular fractionation experiments indicate that there is an increase in both Golgi and cell surface-associated GT pools upon serum-stimulation of resting cells. We conclude that GT is a member of the cell-cycle dependent genes whose expression is growth regulated.