Abstract
The human histone H3 variant, CENP-A, replaces the conventional histone H3 in centromeric chromatin and, together with centromere-specific DNA-binding factors, directs the assembly of the kinetochore. We purified the prenucelosomal e-CENP-A complex. We found that HJURP, a member of the complex, was required for cell cycle specific targeting of CENP-A to centromeres. HJURP facilitated efficient deposition of CENP-A/H4 tetramers to naked DNA in vitro. Bacterially expressed HJURP binds at a stoichiometric ratio to the CENP-A/H4 tetramer but not to the H3/H4 tetramer. The binding occurred through a conserved HJURP short N-terminal domain, termed CBD. The novel characteristic identified in vertebrates that we named TLTY box of CBD, was essential for formation of the HJURP-CENP-A/H4 complex. Our data identified HJURP as a vertebrate CENP-A chaperone and dissected its mode of interactions with CENP-A.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Autoantigens / chemistry*
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Autoantigens / genetics
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Autoantigens / metabolism*
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Cell Cycle
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Centromere / metabolism*
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Centromere Protein A
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Chromosomal Proteins, Non-Histone / chemistry*
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Chromosomal Proteins, Non-Histone / genetics
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Chromosomal Proteins, Non-Histone / metabolism*
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Conserved Sequence
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DNA-Binding Proteins / chemistry*
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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HeLa Cells
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Histones / metabolism
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Humans
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Models, Molecular
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Molecular Sequence Data
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Nucleosomes / metabolism
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Protein Binding
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Protein Interaction Domains and Motifs*
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Protein Multimerization
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Protein Structure, Quaternary
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RNA, Small Interfering / genetics
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Sequence Alignment
Substances
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Autoantigens
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CENPA protein, human
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Centromere Protein A
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Chromosomal Proteins, Non-Histone
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DNA-Binding Proteins
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HJURP protein, human
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Histones
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Nucleosomes
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RNA, Small Interfering