title: Targeting of Arabidopsis KNL2 to Centromeres Depends on the Conserved CENPC-k Motif in Its C Terminus.
abstract: KINETOCHORE NULL2 (KNL2) is involved in recognition of centromeres and in centromeric localization of the centromere-specific histone cenH3. Our study revealed a cenH3 nucleosome binding CENPC-k motif at the C terminus ofArabidopsis thalianaKNL2, which is conserved among a wide spectrum of eukaryotes. Centromeric localization of KNL2 is abolished by deletion of the CENPC-k motif and by mutating single conserved amino acids, but can be restored by insertion of the corresponding motif of Arabidopsis CENP-C. We showed by electrophoretic mobility shift assay that the C terminus of KNL2 binds DNA sequence-independently and interacts with the centromeric transcripts in vitro. Chromatin immunoprecipitation with anti-KNL2 antibodies indicated that in vivo KNL2 is preferentially associated with the centromeric repeatpAL1Complete deletion of the CENPC-k motif did not influence its ability to interact with DNA in vitro. Therefore, we suggest that KNL2 recognizes centromeric nucleosomes, similar to CENP-C, via the CENPC-k motif and binds adjoining DNA.
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, D-06466 Stadt Seeland, Germany.
authors.forename: Michael
authors.identifier: 0000-0001-6893-8431
authors.initials: M
authors.lastname: Sandmann
authors.affiliation: Howard Hughes Medical Institute, Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109.
authors.forename: Paul
authors.initials: P
authors.lastname: Talbert
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, D-06466 Stadt Seeland, Germany.
authors.forename: Dmitri
authors.initials: D
authors.lastname: Demidov
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, D-06466 Stadt Seeland, Germany.
authors.forename: Markus
authors.identifier: 0000-0003-3104-0825
authors.initials: M
authors.lastname: Kuhlmann
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, D-06466 Stadt Seeland, Germany.
authors.forename: Twan
authors.identifier: 0000-0001-5891-6503
authors.initials: T
authors.lastname: Rutten
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, D-06466 Stadt Seeland, Germany.
authors.forename: Udo
authors.identifier: 0000-0002-9416-3470
authors.initials: U
authors.lastname: Conrad
authors.affiliation: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben, D-06466 Stadt Seeland, Germany lermonto@ipk-gatersleben.de.
authors.forename: Inna
authors.identifier: 0000-0003-3386-2590
authors.initials: I
authors.lastname: Lermontova
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002503:Centromere / Q000235:genetics / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D018744:DNA, Plant / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D018386:Kinetochores / Q000378:metabolism; D008957:Models, Genetic; D009154:Mutation; D009707:Nucleosomes / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011485:Protein Binding
chemical_list: D029681:Arabidopsis Proteins; D002868:Chromosomal Proteins, Non-Histone; D018744:DNA, Plant; D004268:DNA-Binding Proteins; C000624953:KNL2 protein, Arabidopsis; D009707:Nucleosomes; C060905:centromere protein C