Knob Heterochromatin Research Articles

Highly repeated DNA sequence limited to knob heterochromatin in maize

A highly repeated DNA sequence has been isolated from the maize genome as a satellite in actinomycin D/CsCl gradients. By using maize stocks differing in their heterochromatin content we have established that the sequence is a major constituent of one class of heterochromatin, knob heterochromatin, which can occur at 23 locations in the chromosome complement. The repeating unit, of 185 base pairs, has been cloned in plasmid pBR322 and its nucleotide sequence has been determined. The presence of this DNA sequence in knob heterochromatin and its absence from centromeric, nucleolar, and B chromosome heterochromatin parallels the cytogenetic differentiation previously described for these classes of heterochromatin in maize. Because knob heterochromatin has a distinctive cytological appearance and is unique in showing neocentric activity at meiosis, its association with a particular repeated DNA sequence may reflect a functional role for the sequence in the cell cycle.

Asynchronous replication of heterochromatin in maize

Replication patterns of four classes of heterochromatin in the chromosomes of maize were analyzed. Centromeric heterochromatin, distal heterochromatin of the B chromosomes, and knob heterochromatin all have asynchronous late replication, knob heterochromatin being the last chromosomal DNA to complete replication. Nucleolus organizer heterochromatin replicates during the period of euchromatin DNA replication. The proportion of the S period involved in asynchronous late replication is directly dependent on the amount of heterochromatin in the nucleus. Both additional knob and B-chromosome heterochromatin participate in this effect. Each additional B chromosome produces an increment in the proportion of nuclei with asynchronous replication; in nuclei with 12 B chromosomes, the proportion of the S phase showing asynchronous late replication is more than 3 times greater than that in nuclei without B chromosomes. The data are consistent with the hypothesis that delayed DNA replication of knob heterochromatin is responsible, in some genotypes, for the loss of chromosome segments and for B-chromosome nondisjunction in the second pollen grain mitosis.