The nuclear location and relocation of genes play crucial regulatory roles in gene expression. SATB1, a MAR-binding protein, has been found to regulate beta-like globin genes through chromatin remodeling. In this study, we generated K562 cells over-expressing wild-type or nuclear matrix targeting sequences (NMTS)-deficient SATB1 and found that like wild-type SATB1, NMTS-deficient SATB1 induces out loop of beta-globin cluster from its chromosome territory (CT), while it is unable to associate the Cluster with the nuclear matrix as wild-type SATB1 does and had no regulatory functions to the beta-globin cluster. Besides, Our data showed that the transacting factor occupancies and chromatin modifications at beta-globin Cluster were differentially affected by wild-type and NMTS-deficient SATB1. These results indicate that SATB1 regulates beta-like globin genes at the nuclear level interlaced with chromatin and DNA level, and emphasize the nuclear matrix binding activity of SATB1 to its regulatory function. (C) 2009 Elsevier Inc. All rights reserved.