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PMID: 2364428 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A programmed site-specific DNA rearrangement in Tetrahymena thermophila requires flanking polypurine tracts.

Cell ·Vol. 61 ·No. 7 ·1990-06-29 ·Pages 1237-46

Godiska R, Yao MC

Abstract

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.

MeSH Terms
Animals Base Sequence Cell Line Chromosome Deletion Cloning, Molecular DNA/genetics Gene Rearrangement Genetic Vectors Molecular Sequence Data Mutation Oligonucleotide Probes Plasmids Repetitive Sequences, Nucleic Acid Tetrahymena/genetics
Chemicals
Oligonucleotide Probes DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Godiska R
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Yao M C
Article Info
Journal
Cell
Abbr.
Cell
ISSN
0092-8674
Published
1990-06-29
Pages
1237-46
Language
English
Region
United States
NLM ID
0413066
Subset
IM
Grants
NIGMS NIH HHS · GM26210 · United States
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