In Saccharomyces cerevisiae, subtelomeric silencing is involved in the propagation of Silent Information Regulator (SIR) proteins toward euchromatin. Numerous mechanisms are involved in antagonizing the local spread of Sir-dependent silent chromatin into neighboring euchromatin. Here, we identified a novel role for sumoylation E3 ligase Mms21 in the maintenance of subtelomeric silencing. We found that disruption of E3 ligase activity of Mms21 results in the de-repression of subtelomeric silencing. Deletion of E3 ligase domain of Mms21 led to decreased binding of Sir2p, Sir3p and Sir4 at subtelomeric chromatins and increased H3K4 tri-methylation at telomere-distal euchromatin regions, correlating with increased gene expression in two subtelomeric reporter genes. In addition, a mms21 Delta sl mutant caused a severe growth defect in combination with htz1 Delta deletion and showed an enhanced association of Htz1 with telomere proximal regions. Taken together, our findings suggest an important role of Mms21p; it contributes to subtelomeric silencing during the formation of a heterochromatin boundary. (C) 2013 Elsevier Inc. All rights reserved.