Journal
EMBO JOURNAL
Volume 20, Issue 12, Pages 3251-3261Publisher
OXFORD UNIV PRESS
DOI: 10.1093/emboj/20.12.3251
Keywords
ciliated protozoa; developmentally programmed genome rearrangements; DNA cleavage; DNA recombination; IES element
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DNA sequences (IES elements) eliminated from the developing macronucleus in the ciliate Tetrahymena thermophila are released as linear fragments, which have now been detected and isolated. A PCR-mediated examination of fragment end structures reveals three types of strand scission events, reflecting three steps in the deletion process. New evidence is provided for two steps proposed previously: an initiating double-stranded cleavage, and strand transfer to create a branched deletion intermediate. The fragment ends provide evidence for a previously uncharacterized third step: the branched DNA strand is cleaved at one of several defined sites located within 15-16 nucleotides of the IES boundary, liberating the deleted DNA in a linear form.
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