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Modular structural elements in the replication origin region of Tetrahymena rDNA
C Du1, R P Sanzgiri, W L Shaiu
1Department of Zoology and Genetics, Iowa State University, Ames 50011, USA.
Nucleic Acids Research
|May 25, 1995
Summary
Computer analyses identified a DNA replication origin in Tetrahymena thermophila. This protozoan origin shares conserved structural elements with eukaryotic chromosomal origins, suggesting a common evolutionary feature.
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- DNA replication origins are crucial for cell division.
- Eukaryotic origins often contain conserved modular sequence elements.
- The Tetrahymena thermophila rRNA gene 5'NTS was a candidate replication origin.
Purpose of the Study:
- To investigate the DNA replication origin in Tetrahymena thermophila.
- To identify structural elements within the 5'NTS region.
- To determine if these elements are conserved across eukaryotes.
Main Methods:
- Computer analysis of DNA sequences.
- Mung bean nuclease sensitivity assays.
- Electrophoresis to detect bent DNA.
- In vitro nuclear matrix binding assays.
Main Results:
- Identified a potential DNA unwinding element (DUE) and bent DNA segments in the 5'NTS.
- Localized hypersensitive sites within the DUE-like element.
- Confirmed association of the 5'NTS with the nuclear matrix in vitro.
Conclusions:
- The Tetrahymena 5'NTS contains structural features characteristic of eukaryotic replication origins.
- These findings suggest conserved modular elements at eukaryotic chromosomal origins.
- This study provides direct evidence in a protozoan origin region.