Differential response of the human cyclin B1 promoter to inhibitors of the cell cycle in NIH3T3 cells

In this study, NIH3T3 cells stably transfected with a cyclin B1-luciferase reporter vector were utilized to investigate if cyclin B1 promoter activity is linked to either DNA replication or the activities of various cyclin-cyclin dependent kinases (cdks). Synchronized cells treated at the time of serum re-stimulation with 2 mu g/ml of the DNA synthesis inhibitor, aphidicolin, did not display an increase in luciferase activity in comparison to control cells. When treated with aphidicolin during S phase, luciferase activity decreased. In contrast, luciferase activity increased in cells treated at the time of serum re-stimulation with 200 mu M olomoucine, a cyclin-cdk inhibitor. These results indicate that (1) cyclin B1 promoter activity in NIH3T3 cells is linked to a DNA replication checkpoint control mechanism; (2) the cyclin B1 gene can be activated in the absence of functional cyclin E-cdk2, cyclin A-cdk2, or cyclin B-cdk2; and (3) cyclin B1 gene activation can occur in G1 arrested cells under conditions in which the arrest is not directly linked to inhibition of DNA synthesis. (C) 1998 Academic Press.