Creatine kinase-B (CKB) synthesis is rapidly and specifically induced by estrogen in the uterus of the immature rat. This study indicates that this elevation is due at least in part to increases in the levels of mRNA for CKB. The stimulation of CKB mRNA levels is rapid (a 7- to 10-fold increase is detected 1-3 h after estrogen administration), but transient, as levels return to near control values by 6 h. Analysis of cDNAs to both uterine and brain CKB mRNA indicate that the same sequence is expressed in both tissues despite earlier observations of heterogeneity of the protein isolated from the two tissues. A 1.7-kilobasepair DNA fragment containing the CKB promoter and 5' flanking sequences confers estrogen sensitivity on expression of the bacterial chloramphenicol acetyl transferase gene in HeLa cells on cotransfection with an estrogen-receptor expression vector. However, the CKB promoter sequences lack any motif with convincing similarity to the currently accepted consensus estrogen response element GGTCAnnnTGACC.