A programmed site-specific DNA rearrangement in Tetrahymena thermophila requires flanking polypurine tracts.
During macronuclear development in ciliates, precise deletion events eliminate thousands of specific DNA segments. Each segment is bounded by a unique pair of short direct repeats, but no other common feature has been reported. To determine the critical cis-acting sequences, we developed an in vivo system for analyzing this process in Tetrahymena. We show that sequences essential for recognition and excision of one such region are located within the 70 bp of DNA flanking either side of it. Three authentic splice sites and one cryptic site are each adjacent to an unusual polypurine tract (5'-A5G5) situated 40-50 bp distal to each terminal repeat. Removal of this tract or substitution of 3 bp within it abolishes splicing to the adjacent site. The normal chromosomal environment and the integrity of the eliminated sequence are not required for its removal. We believe the polypurine tract is a signal essential for excision of this sequence.
Godiska
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R
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Yao
M C
MC
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D015321:Gene Rearrangement; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D010957:Plasmids; D012091:Repetitive Sequences, Nucleic Acid; D013768:Tetrahymena / Q000235:genetics*
D015345:Oligonucleotide Probes; D004247:DNA