title: Putative CENP-B paralogues are not present at mammalian centromeres.
abstract: Although centromere protein B (CENP-B) is a highly conserved mammalian centromere protein, its function remains unknown. The presence of the protein is required to form artificial satellite DNA-based centromeres de novo, yet cenpb knockout mice are viable for multiple generations with no mitotic or meiotic defects, and the protein is not present at fully functional neocentromeres. Previous studies have suggested that the presence of functionally redundant paralogues of CENP-B may explain the lack of a phenotype in knockout mice, and the related Tigger-derived (TIGD) family of proteins has been implicated as the most likely candidate for such paralogues. Here, we describe an investigation of the centromere-binding properties of the three TIGD proteins most highly related to CENP-B through phylogenetic analysis through EGFP fusion studies and immunocytochemistry. Although two of the three proteins bound to human centromeres with low affinity when overexpressed as fusion proteins, the strongest candidate, TIGD3, demonstrated no native centromeric binding when using raised antibodies, either in human cells or in cenpb                         (-/-) mouse ES cells. We conclude that the existence of functional CENP-B paralogues is highly unlikely and that CENP-B acts alone at the centromere. Based on these data, we suggest a new, meiotic drive model of CENP-B action during centromere repositioning in evolution.
authors.affiliation: Chromosome and Chromatin Research, Murdoch Childrens Research Institute, Royal Children's Hospital, Parkville, Australia. owen.marshall@mcri.edu.au
authors.forename: Owen J
authors.initials: OJ
authors.lastname: Marshall
authors.forename: K H Andy
authors.initials: KH
authors.lastname: Choo
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002503:Centromere / Q000235:genetics*; D051936:Centromere Protein B / Q000235:genetics*; D003001:Cloning, Molecular; D019295:Computational Biology; D017931:DNA Primers / Q000235:genetics; D019143:Evolution, Molecular; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D018345:Mice, Knockout; D005810:Multigene Family / Q000235:genetics*; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D051936:Centromere Protein B; D017931:DNA Primers; D009687:Nuclear Proteins; C573495:TIGD3 protein, human; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
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