Phosphorylation of keratin polypeptides was studied by incubating vaginal tissues (removed from estradiol primed and unprimed 30-day-old rats) with 32Pi. Analysis by SDS-PAGE and autoradiography showed that on treatment with estradiol phosphorylation of 63 and 58 kDa keratin polypeptides increased 3- and 2-fold respectively. Phosphorylation was maximal after 30 min of estradiol priming and decreased thereafter. Phosphorylation of some non-keratin polypeptides (37, 34, 32 and 25 kDa) also showed time dependent variation. The results showed that estradiol can modulate phosphorylation-dephosphorylation of keratins and other polypeptides in rat vaginal epithelial cells.