title: The three methyl-CpG-binding domains of AtMBD7 control its subnuclear localization and mobility.
abstract: Three methyl-CpG-binding domain (MBD) proteins in Arabidopsis, AtMBD5, AtMBD6, and AtMBD7, are functional in binding methylated CpG dinucleotides in vitro and localize to the highly CpG-methylated chromocenters in vivo. These proteins differ, however, in their subnuclear localization pattern; AtMBD5 and AtMBD6, each containing a single MBD motif, show preference for two perinucleolar chromocenters, whereas AtMBD7, a naturally occurring poly-MBD protein containing three MBD motifs, localizes to all chromocenters. Here we studied the significance of multiple MBD motifs for subnuclear localization and mobility in living cells. We found that the number of MBD motifs determines the subnuclear localization of the MBD protein. Furthermore, live kinetic experiments showed that AtMBD7-green fluorescent protein (GFP) has lower mobility than AtMBD5-GFP and AtMBD6-GFP, which is conferred by cooperative activity of its three MBD motifs. Thus, the number of MBD motifs appears to affect not only binding affinity and mobility within the nucleus, but also the subnuclear localization of the protein. Our results suggest that poly-MBD proteins can directly affect chromatin structure by inducing intra- and inter-chromatin compaction via bridging over multiple methylated CpG sites.
authors.affiliation: Department of Plant Sciences, The Weizmann Institute of Science, Rehovot, Israel. assaf.zemach@weizmann.ac.il
authors.forename: Assaf
authors.initials: A
authors.lastname: Zemach
authors.forename: Ofer
authors.initials: O
authors.lastname: Gaspan
authors.forename: Gideon
authors.initials: G
authors.lastname: Grafi
mesh_terms: D020816:Amino Acid Motifs; D017360:Arabidopsis; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D018899:CpG Islands / Q000235:genetics*; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; D004268:DNA-Binding Proteins; C525054:MBD7 protein, Arabidopsis