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Inter-species DNA polymerase delta chimeras are functional in Saccharomyces cerevisiae
G Moussy1, A M De Recondo, G Baldacci
1Institut de Recherches sur le Cancer, Centre National de la Recherche Scientifique Unité 9044, Villejuif, France.
European Journal of Biochemistry
|July 1, 1995
Summary
Investigating DNA polymerase delta in yeast species revealed species-specific functional differences. A specific 3' region of the Saccharomyces cerevisiae POL3 gene dictates these species-specific interactions, impacting DNA replication and repair.
Area of Science:
- Molecular Biology
- Yeast Genetics
- DNA Replication
Background:
- High sequence identity (>50%) exists between the catalytic subunits of DNA polymerase delta in Schizosaccharomyces pombe and Saccharomyces cerevisiae.
- Understanding species-specific functional divergence in essential proteins like DNA polymerase delta is crucial for evolutionary and functional genomics.
Purpose of the Study:
- To investigate the functional complementation capabilities of Schizosaccharomyces pombe DNA polymerase delta in Saccharomyces cerevisiae.
- To identify specific regions within the POL3 gene responsible for species-specific interactions and functions of DNA polymerase delta.
Main Methods:
- In vivo complementation assays using thermosensitive mutants of Saccharomyces cerevisiae.
- Construction and functional analysis of chimeric DNA polymerase delta genes between S. pombe and S. cerevisiae.
- Assessing the growth of a S. cerevisiae strain with a disrupted POL3 gene after transformation with chimeric constructs.
Main Results:
- Schizosaccharomyces pombe DNA polymerase delta showed allele-dependent complementation of S. cerevisiae thermosensitive mutants.
- S. pombe DNA polymerase delta failed to restore growth in a S. cerevisiae strain with a disrupted POL3 gene.
- A 1254-bp region in the 3' end of the Saccharomyces cerevisiae POL3 gene was identified as responsible for species-specific functions.
Conclusions:
- Functional differences exist between yeast DNA polymerase delta proteins from different species, despite high sequence homology.
- A defined region in the 3' portion of the Saccharomyces cerevisiae POL3 gene confers species-specific functional properties to DNA polymerase delta.