Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
???displayArticle.abstract???
We have taken advantage of the known structural parameters associated with centromere DNA in vivo to construct a CEN fragment that can be selectively excised from the chromatin DNA with restriction endonucleases. CEN3 DNA is organized in chromatin such that a 220-250-bp region encompassing the elements of centromere homology is resistant to nuclease digestion. Restriction enzyme linkers encoding the Bam HI-recognition site were ligated to a 289 base pair DNA segment that spans the 220-250-bp protected core (Bloom et al., 1984). Replacement of this CEN3-Bam HI linker cassette into a chromosome or plasmid results in formation of a complete structural and functional centromeric unit. A centromere core complex that retains its protected chromatin conformation can be selectively excised from intact nuclei by restriction with the enzyme Bam HI. The centromeric protein-DNA complex is therefore not dependent upon the intact torsional constrains on linear chromosomes for its structural integrity. Isolation of this complex provides a novel approach to characterizing authentic centromeric proteins bound to DNA in their native state.
Bloom,
Yeast centromere DNA is in a unique and highly ordered structure in chromosomes and small circular minichromosomes.
1982, Pubmed
Bloom,
Yeast centromere DNA is in a unique and highly ordered structure in chromosomes and small circular minichromosomes.
1982,
Pubmed
Bloom,
Chromatin conformation of yeast centromeres.
1984,
Pubmed
,
Xenbase
Bloom,
Fractionation of hen oviduct chromatin into transcriptionally active and inactive regions after selective micrococcal nuclease digestion.
1978,
Pubmed
Carbon,
Structural and functional analysis of a yeast centromere (CEN3).
1984,
Pubmed
Fitzgerald-Hayes,
Nucleotide sequence comparisons and functional analysis of yeast centromere DNAs.
1982,
Pubmed
Forte,
Naturally occurring cross-links in yeast chromosomal DNA.
1976,
Pubmed
Gaudet,
Alterations in the adenine-plus-thymine-rich region of CEN3 affect centromere function in Saccharomyces cerevisiae.
1987,
Pubmed
Hegemann,
Mutations in the right boundary of Saccharomyces cerevisiae centromere 6 lead to nonfunctional or partially functional centromeres.
1986,
Pubmed
Hill,
Genetic manipulation of centromere function.
1987,
Pubmed
Karin,
Primary structure and transcription of an amplified genetic locus: the CUP1 locus of yeast.
1984,
Pubmed
Larsen,
An altered DNA conformation detected by S1 nuclease occurs at specific regions in active chick globin chromatin.
1982,
Pubmed
Lilley,
The inverted repeat as a recognizable structural feature in supercoiled DNA molecules.
1980,
Pubmed
McGhee,
A 200 base pair region at the 5' end of the chicken adult beta-globin gene is accessible to nuclease digestion.
1981,
Pubmed
McGrew,
Single base-pair mutations in centromere element III cause aberrant chromosome segregation in Saccharomyces cerevisiae.
1986,
Pubmed
Nelson,
Nucleosome organization of the yeast 2-micrometer DNA plasmid: a eukaryotic minichromosome.
1979,
Pubmed
Panzeri,
Role of conserved sequence elements in yeast centromere DNA.
1985,
Pubmed
Rothstein,
One-step gene disruption in yeast.
1983,
Pubmed
Saunders,
Chromatin structure of altered yeast centromeres.
1988,
Pubmed
Smart,
Selective dissociation of histones from chromatin by sodium deoxycholate.
1971,
Pubmed
Southern,
Detection of specific sequences among DNA fragments separated by gel electrophoresis.
1975,
Pubmed
Thomas,
Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.
1980,
Pubmed
Weintraub,
A dominant role for DNA secondary structure in forming hypersensitive structures in chromatin.
1983,
Pubmed
