Localisation of centromeric proteins to a fraction of mouse minor satellite DNA on a mini-chromosome in human, mouse and chicken cells.
Centromeres are required for faithful segregation of chromosomes in cell division. It is not clear how centromere sites are specified on chromosomes in vertebrates. We have previously introduced a mini-chromosome, named ST1, into a variety of cell lines including human HT1080, mouse LA9 and chicken DT40. This mini-chromosome, segregating faithfully in these cells, contains mouse minor and major, and human Y alpha-satellite DNA repeats. In this study, after determining the organisation of the satellite repeats, we investigated the location of the centromere on the mini-chromosome by combined immunocytochemistry and fluorescence in situ hybridisation analysis. Centromeric proteins were consistently co-localised with the minor satellite repeats in all three cell lines. When chromatin fibres were highly stretched, centromeric proteins were only seen on a small portion of the minor satellite repeats. These results indicate that a fraction of the minor satellite repeats is competent in centromere function not only in mouse but also in human and chicken cells.
MRC Human Genetics Unit, Western General Hospital, Crewe Road, Edinburgh, EH4 2XU, UK.
Kang
K
Zeng
Jose I
JI
de las Heras
Andrew
A
Ross
Jian
J
Yang
Howard
H
Cooke
Ming Hong
MH
Shen
D000818:Animals; D002460:Cell Line; D002503:Centromere / Q000235:genetics / Q000502:physiology; D002645:Chickens; D002843:Chromatin / Q000235:genetics / Q000502:physiology; D022221:Chromosomes, Artificial, Mammalian / Q000235:genetics / Q000502:physiology*; D004276:DNA, Satellite / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D051379:Mice; D018598:Minisatellite Repeats / Q000235:genetics / Q000502:physiology*
D002843:Chromatin; D004276:DNA, Satellite; D004268:DNA-Binding Proteins