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Replication forks pause at yeast centromeres.

S A Greenfeder¹, C S Newlon

¹Department of Microbiology and Molecular Genetics, UMDNJ-New Jersey Medical School, Newark 07103.

Molecular and Cellular Biology

|September 1, 1992

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Replication forks pause at yeast centromeres due to protein-DNA complexes, not DNA sequence. This complex, essential for centromere function, is present during DNA replication (S phase).

Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

Yeast centromeric DNA (120 bp) forms a protein-DNA complex, creating a nuclease-resistant core (200-240 bp).
Understanding centromere replication is crucial for chromosome stability and cell division.

Purpose of the Study:

To investigate replication fork behavior at yeast centromeres.
To determine if replication fork pausing is caused by DNA sequence or protein-DNA complex.

Main Methods:

Two-dimensional agarose gel electrophoresis was used to analyze replication of chromosomal and plasmid-based yeast centromeres (CEN1, CEN3, CEN4).
Analysis included mutant centromere derivatives with varying function and core structure formation.

Main Results:

Related Experiment Videos

Replication fork pause sites were identified at CEN1, CEN3, and CEN4, suggesting a general phenomenon at yeast centromeres.
The ability of centromere derivatives to cause replication fork pausing correlated with their ability to form the nuclease-resistant core structure.
Pausing was independent of specific DNA sequences within the centromere.

Conclusions:

The centromere protein-DNA complex, not the DNA sequence itself, is responsible for replication fork pausing.
This protein-DNA complex is likely present throughout the cell cycle, including during S phase when replication occurs.