ArticleChromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology1998
Nucleotide specificity at the boundary and size requirement of the target sites recognized by human centromere protein B (CENP-B) in vitro.
Article in Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology, 1998. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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5 citing papers in PubMed.
- The possible functions of duplicated ets (GGAA) motifs located near transcription start sites of various human genes.Cellular and molecular life sciences : CMLS · 2011Article
- Tandemly repeated DNA sequences and centromeric chromosomal regions of Arabidopsis species.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2003Review
- A tandem repetitive sequence located in the centromeric region of common wheat (Triticum aestivum) chromosomes.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2001Article
- Polymorphisms and genomic organization of repetitive DNA from centromeric regions of Arabidopsis chromosomes.The Plant cell · 1999Article
- Visualization of prekinetochore locus on the centromeric region of highly extended chromatin fibers: does kinetochore autoantigen CENP-C constitute a kinetochore organizing center?Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 1999Article
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Abstract
Human centromere protein B (CENP-B) has a sequence-specific DNA binding activity. We previously reported several CENP-B binding motifs by analysing synthetic oligonucleotides as well as alphoid DNA isolated from the human genomic library. Here, we examined the size requirement and nucleotide specificity of human CENP-B binding sequences in vitro. We synthesized three sets of mixed oligonucleotides containing diverged authentic binding sites (CTTCGTTGGAAACGGGA) in which certain pairs of nucleotides (underlined) were degenerated. Each oligonucleotide with a defined sequence was separately introduced into a plasmid and mixed with GST-fused recombinant CENP-B. The DNA-protein complex formed was affinity purified with glutathione Sepharose. Any nucleotide substitutions at the positions 1, 2 and 17 did not significantly influence the recovery, while the substitutions at positions 3, 4 and 16 did, suggesting that the internal 14-bp motif (TCGTTGGAAACGGG) constituted the minimum requirement. However, it showed a lower affinity to CENP-B, compared with the authentic motif. The inclusion of T at the 5' end greatly increased the affinity, and the further addition of A or T at the 3' end (TTCGTTGGAAACGGGA/T) offered affinity similar to the authentic motif. The first nucleotide of the 17-bp authentic binding motif may not be essential for CENP-B binding.
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