Molecular characterization of human minichromosomes with centromere from chromosome 1 in human-hamster hybrid cells

Abstract

In this study we examine the amounts of four different human satellite DNA sequences in a series of human-hamster hybrid cells, which contain a human minichromosome including the centromere of human chromosome 1. Comparisons with the corresponding amounts in an intact human chromosome 1 suggest that the minichromosomes have lost satellite DNA sequences, and in one case a substantial fraction of several satellite DNAs is lost, without affecting the stability and normal mitotic segregation of the minichromosome. The smallest minichromosome appears to have lost all of the long arm and a significant portion of centromeric heterochromatin, while retaining 1000-2000 kb of the short arm of human chromosome 1. The satellite sequences examined include: a chromosome 1-specific satellite III probe, a chromosome 1-specific alpha satellite DNA, another alpha satellite DNA originally derived from the X chromosome, and an alphoid EcoRI dimer whose isolation from one of the minichromosomes and characterization is also described in this paper. One interpretation of these data indicates that an interspersion of blocks of satellite sequences occurs in the centromere region of chromosome 1. If these satellite sequences have functional significance, then there may be redundancy in the system that allows for a variation in the size of the kinetochore and the number of attachment sites for microtubules.