A conserved nucleotide sequence at the sites of developmentally regulated chromosomal breakage in Tetrahymena

Abstract

Chromosomal breakage occurs at hundreds of specific sites in Tetrahymena, including the two ends of the unique ribosomal RNA genes, during the development of the somatic macronucleus. We have identified a 15-nucleotide sequence that occurs widely in the germinal micronuclear genome and is associated exclusively with chromosomal breakage sites. When copies of this sequence were cloned and analyzed, they were found in all cases to be located at or very near sites of breakage. This general rule is further supported by the observation that in a different site in which a single nucleotide substitution is found within this sequence, no chromosomal breakage occurs. The complete sequence structure of one of the breakage junctions has also been determined in both the germinal DNA and the two somatic DNA termini. This structure reveals that the 15-nucleotide conserved sequence is located within a 54-nucleotide region that is removed following chromosomal breakage.