title: The RPT2 subunit of the 26S proteasome directs complex assembly, histone dynamics, and gametophyte and sporophyte development in Arabidopsis.
abstract: The regulatory particle (RP) of the 26S proteasome contains a heterohexameric ring of AAA-ATPases (RPT1-6) that unfolds and inserts substrates into the core protease (CP) for degradation. Through genetic analysis of the Arabidopsis thaliana gene pair encoding RPT2, we show that this subunit plays a critical role in 26S proteasome assembly, histone dynamics, and plant development. rpt2a rpt2b double null mutants are blocked in both male and female gamete transmission, demonstrating that the subunit is essential. Whereas rpt2b mutants are phenotypically normal, rpt2a mutants display a range of defects, including impaired leaf, root, trichome, and pollen development, delayed flowering, stem fasciation, hypersensitivity to mitomycin C and amino acid analogs, hyposensitivity to the proteasome inhibitor MG132, and decreased 26S complex stability. The rpt2a phenotype can be rescued by both RPT2a and RPT2b, indicative of functional redundancy, but not by RPT2a mutants altered in ATP binding/hydrolysis or missing the C-terminal hydrophobic sequence that docks the RPT ring onto the CP. Many rpt2a phenotypes are shared with mutants lacking the chromatin assembly factor complex CAF1. Like caf1 mutants, plants missing RPT2a or reduced in other RP subunits contain less histones, thus implicating RPT2 specifically, and the 26S proteasome generally, in plant nucleosome assembly.
authors.affiliation: Department of Genetics, University of Wisconsin, Madison, Wisconsin 53706, USA.
authors.forename: Kwang-Hee
authors.initials: KH
authors.lastname: Lee
authors.forename: Atsushi
authors.initials: A
authors.lastname: Minami
authors.forename: Richard S
authors.initials: RS
authors.lastname: Marshall
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Book
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Farmer
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Walker
authors.forename: Richard D
authors.initials: RD
authors.lastname: Vierstra
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000483:Alleles; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002458:Cell Fractionation; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D005816:Genetic Complementation Test; D056426:Genetic Loci; D055993:Germ Cells, Plant / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism; D015151:Immunoblotting; D016685:Mitomycin / Q000494:pharmacology; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010641:Phenotype; D018517:Plant Roots / Q000254:growth & development / Q000378:metabolism; D011058:Pollen / Q000235:genetics / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D015398:Signal Transduction; D019076:Transgenes
chemical_list: D029681:Arabidopsis Proteins; D006657:Histones; C487429:RPT2a protein, Arabidopsis; D016685:Mitomycin; D000255:Adenosine Triphosphate; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease