title: N terminus of Swr1 binds to histone H2AZ and provides a platform for subunit assembly in the chromatin remodeling complex.
abstract: Variant histone H2AZ-containing nucleosomes are involved in the regulation of gene expression. In Saccharomyces cerevisiae, chromatin deposition of histone H2AZ is mediated by the fourteen-subunit SWR1 complex, which catalyzes ATP-dependent exchange of nucleosomal histone H2A for H2AZ. Previous work defined the role of seven SWR1 subunits (Swr1 ATPase, Swc2, Swc3, Arp6, Swc5, Yaf9, and Swc6) in maintaining complex integrity and H2AZ histone replacement activity. Here we examined the function of three additional SWR1 subunits, bromodomain containing Bdf1, actin-related protein Arp4 and Swc7, by analyzing affinity-purified mutant SWR1 complexes. We observed that depletion of Arp4 (arp4-td) substantially impaired the association of Bdf1, Yaf9, and Swc4. In contrast, loss of either Bdf1 or Swc7 had minimal effects on overall complex integrity. Furthermore, the basic H2AZ histone replacement activity of SWR1 in vitro required Arp4, but not Bdf1 or Swc7. Thus, three out of fourteen SWR1 subunits, Bdf1, Swc7, and previously noted Swc3, appear to have roles auxiliary to the basic histone replacement activity. The N-terminal region of the Swr1 ATPase subunit is necessary and sufficient to direct association of Bdf1 and Swc7, as well as Arp4, Act1, Yaf9 and Swc4. This same region contains an additional H2AZ-H2B specific binding site, distinct from the previously identified Swc2 subunit. These findings suggest that one SWR1 enzyme might be capable of binding two H2AZ-H2B dimers, and provide further insight on the hierarchy and interdependency of molecular interactions within the SWR1 complex.
authors.affiliation: Laboratory of Biochemistry and Molecular Biology, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA. wuwe@mail.nih.gov
authors.forename: Wei-Hua
authors.initials: WH
authors.lastname: Wu
authors.forename: Chwen-Huey
authors.initials: CH
authors.lastname: Wu
authors.forename: Andreas
authors.initials: A
authors.lastname: Ladurner
authors.forename: Gaku
authors.initials: G
authors.lastname: Mizuguchi
authors.forename: Debbie
authors.initials: D
authors.lastname: Wei
authors.forename: Hua
authors.initials: H
authors.lastname: Xiao
authors.forename: Ed
authors.initials: E
authors.lastname: Luk
authors.forename: Anand
authors.initials: A
authors.lastname: Ranjan
authors.forename: Carl
authors.initials: C
authors.lastname: Wu
mesh_terms: D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism; D000199:Actins / Q000235:genetics / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly / Q000502:physiology*; D051548:Histone Acetyltransferases; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D009707:Nucleosomes / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D000092468:Bromodomain Containing Proteins
chemical_list: D000123:Acetyltransferases; D000199:Actins; D000251:Adenosine Triphosphatases; D051548:Histone Acetyltransferases; D006657:Histones; D046912:Multiprotein Complexes; D009687:Nuclear Proteins; D009707:Nucleosomes; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C465210:Act1 protein, S cerevisiae; C088827:Arp4 protein, S cerevisiae; C091342:BDF1 protein, S cerevisiae; D000092468:Bromodomain Containing Proteins; C493530:Htz1 protein, S cerevisiae; C503153:Swc4 protein, S cerevisiae; C477280:YAF9 protein, S cerevisiae; C481261:Swr1 protein, S cerevisiae