NDLI: Centromere protein B assembles human centromeric alpha-satellite DNA at the 17-bp sequence, CENP-B box.

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Centromere protein B assembles human centromeric alpha-satellite DNA at the 17-bp sequence, CENP-B box.

Content Provider	Scilit
Author	Muro, Y. Masumoto, H. Yoda, K. Nozaki, N. Ohashi, M. Okazaki, T.
Copyright Year	1992
Description	We purified 15,000-fold from HeLa cell nuclear extract the centromere antigen that reacts specifically with the 17-bp sequence, designated previously as CENP-B box, in human centromeric alpha-satellite (alphoid) DNA by a two-step procedure including an oligonucleotide affinity column. The purified protein was identified as the centromere protein B (CENP-B) by its mobility on SDS-PAGE (80 kD), and reactivities to a monoclonal antibody raised to CENP-B (bacterial fusion protein) and to anticentromere sera from patients with autoimmune diseases. Direct binding by CENP-B of the CENP-B box sequence in the alphoid DNA has been proved using the purified CENP-B by DNA mobility-shift assay, Southwestern blotting, and DNase I protection analysis. The binding constant of the antigen to the CENP-B box sequence is 6 x 10(8) M-1. DNA mobility-shift assays indicated that the major complex formed between the CENP-B and the DNA contains two DNA molecules, suggesting the importance of the CENP-B/CENP-B box interaction in organization of higher ordered chromatin structures in the centromere and/or kinetochore. Location of DNA binding and dimerization domains in CENP-B was discussed based on the DNA mobility-shift assays performed with a protein fraction containing intact and partial cleavage products of CENP-B.
Related Links	http://jcb.rupress.org/content/jcb/116/3/585.full.pdf https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2289308/pdf
Ending Page	596
Page Count	12
Starting Page	585
DOI	10.1083/jcb.116.3.585
Journal	Journal of Cell Biology
Issue Number	3
Volume Number	116
Language	English
Publisher	Rockefeller University Press
Publisher Date	1992-02-01
Access Restriction	Open
Subject Keyword	Biochemistry and Molecular Biology Dna Binding Proteins Satellite Dna Centromere Journal: Journal of Cell Biology (Vol- 116, Issue- 3)
Content Type	Text

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