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In vitro assembly of the CENP‐B/α‐satellite DNA/core histone complex: CENP‐B causes nucleosome positioning
Author(s) -
Yoda Kinya,
Ando Satoshi,
Okuda Asuko,
Kikuchi Akihiko,
Okazaki Tuneko
Publication year - 1998
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1046/j.1365-2443.1998.00210.x
Subject(s) - nucleosome , biology , centromere , histone , micrococcal nuclease , dna , chromatosome , linker dna , microbiology and biotechnology , biophysics , genetics , chromosome , gene
Background We have studied the nucleosome structure formed from α‐satellite DNA bound with CENP‐B and core histones, in order to develop a previous proposal that the CENP‐B dimer may play a critical role in the assembly of higher order structures of the human centromere by juxtaposing CENP‐B boxes in long α‐satellite arrays. Results The dimeric structure of CENP‐B was sufficiently stable to bundle together two 3.5 kbp DNA fragments when each DNA contained a CENP‐B box. When the same length of DNA included two CENP‐B boxes, the intra–molecular interaction with the CENP‐B dimer predominated, resulting in the formation of loop structures. The in vitro assembly of CENP‐B/α‐satellite DNA/core histone complexes with the aid of nucleosome assembly protein‐1 (NAP‐1) permitted an investigation into the nucleosome arrangement in α‐satellite DNA with CENP‐B bound to CENP‐B boxes. Footprint analyses with micrococcal nuclease (MNase) revealed that CENP‐B causes nucleosome positioning between pairs of CENP‐B boxes with unique hypersensitive sites created on both sides. Conclusion We propose that CENP‐B functions as a structural factor in the centromere region in order to establish a unique, centromere specific pattern of nucleosome positioning.