Silencing of the transcriptional repressor REST is required for terminal differentiation of neuronal and beta-cells. In this study, we hypothesized that REST expression is controlled by hairy and enhancer of split 1 (HES-1), a transcriptional repressor that plays an important role in brain and pancreas development. We identified several N elements (CTNGTG) within the promoter of REST and confirmed that HES-1 associates with the endogenous promoter of REST. Moreover, using a cells model that overexpress HES-1 and a combination of experimental approaches, we demonstrated that HES-1 reduces endogenous REST expression. Taken together, these results indicate that HES-1 is an upstream negative regulator of REST expression.