The initiation of protein synthesis by ribosomal particles of rat ventral prostate was studied by measuring ribosomal binding of an initiator (35-S)methionyl-tRNAf. The binding activity is dependent on ribosomes, GTP, and a prostate cytosol protein fraction. The 40S but not the 60S ribosomal subunit particles are active. The cytosol activity decreases rapidly within one hour after the rat is castrated. This loss is prevented by an intraperitoneal injection of 17beta-hydroxy-5-alpha-androstan-3-one (5-alpha-dihydrotestosterone). The cytosol activity can be stimulated almost immediately (within 10 min) after an intravenous injection of low dose (15 mug per rat) of 5-alpha-dihydrotestosterone into the castrated rat.