title: MAPKAP kinase 3pK phosphorylates and regulates chromatin association of the polycomb group protein Bmi1.
abstract: Polycomb group (PcG) proteins form chromatin-associated, transcriptionally repressive complexes, which are critically involved in the control of cell proliferation and differentiation. Although the mechanisms involved in PcG-mediated repression are beginning to unravel, little is known about the regulation of PcG function. We showed previously that PcG complexes are phosphorylated in vivo, which regulates their association with chromatin. The nature of the responsible PcG kinases remained unknown. Here we present the novel finding that the PcG protein Bmi1 is phosphorylated by 3pK (MAPKAP kinase 3), a convergence point downstream of activated ERK and p38 signaling pathways and implicated in differentiation and developmental processes. We identified 3pK as an interaction partner of PcG proteins, in vitro and in vivo, by yeast two-hybrid interaction and co-immunoprecipitation, respectively. Activation or overexpression of 3pK resulted in phosphorylation of Bmi1 and other PcG members and their dissociation from chromatin. Phosphorylation and subsequent chromatin dissociation of PcG complexes were expected to result in de-repression of targets. One such reported Bmi1 target is the Cdkn2a/INK4A locus. Cells overexpressing 3pK showed PcG complex/chromatin dissociation and concomitant de-repression of p14(ARF), which was encoded by the Cdkn2a/INK4A locus. Thus, 3pK is a candidate regulator of phosphorylation-dependent PcG/chromatin interaction. We speculate that phosphorylation may not only affect chromatin association but, in addition, the function of individual complex members. Our findings linked for the first time MAPK signaling pathways to the Polycomb transcriptional memory system. This suggests a novel mechanism by which a silenced gene status can be modulated and implicates PcG-mediated repression as a dynamically controlled process.
authors.affiliation: Research Institute Growth and Development, Molecular Genetics, Maastricht University, Universiteitssingel 50, 6200 MD, Maastricht, The Netherlands. w.voncken@gen.unimaas.nl
authors.forename: Jan Willem
authors.initials: JW
authors.lastname: Voncken
authors.forename: Hanneke
authors.initials: H
authors.lastname: Niessen
authors.forename: Bernd
authors.initials: B
authors.lastname: Neufeld
authors.forename: Ulrike
authors.initials: U
authors.lastname: Rennefahrt
authors.forename: Vivian
authors.initials: V
authors.lastname: Dahlmans
authors.forename: Nard
authors.initials: N
authors.lastname: Kubben
authors.forename: Barbara
authors.initials: B
authors.lastname: Holzer
authors.forename: Stephan
authors.initials: S
authors.lastname: Ludwig
authors.forename: Ulf R
authors.initials: UR
authors.lastname: Rapp
mesh_terms: D002460:Cell Line; D002843:Chromatin / Q000378:metabolism*; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008934:Mitogens / Q000494:pharmacology; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D063150:Polycomb Repressive Complex 1; D063151:Polycomb Repressive Complex 2; D063146:Polycomb-Group Proteins; D011485:Protein Binding / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013379:Substrate Specificity; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: C069077:BMI1 protein, human; D002843:Chromatin; D019941:Cyclin-Dependent Kinase Inhibitor p16; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D008934:Mitogens; D009687:Nuclear Proteins; C485977:PHC2 protein, human; D063146:Polycomb-Group Proteins; D011518:Proto-Oncogene Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D063151:Polycomb Repressive Complex 2; D063150:Polycomb Repressive Complex 1; C098416:MAP-kinase-activated kinase 3; D017346:Protein Serine-Threonine Kinases