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Yeast mutator phenotype enforced by Arabidopsis PMS1 expression

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Abstract

The DNA mismatch repair (MMR) system is a major DNA repair pathway whose function is critical for the correction of DNA biosynthetic errors. MMR is initiated by the binding of MutS proteins to mismatches and unpaired nucleotides followed by the recruitment of MutL proteins. The major MutL activity in eukaryotes is performed by MutLα, the heterocomplex of MLH1-PMS1 in yeast and plants and MLH1-PMS2 in humans. We here report the effect the expression of Arabidopsis PMS1 protein exerts on Saccharomyces cerevisiae genomic stability. A strain carrying specific microsatellite instability reporter systems was chosen for the study. The plant protein failed to complement the hypermutator phenotype of a pms1 deficient strain but increased approximately 14-fold and 2,000-fold the mutation rates of his7-2 and lys2::InsE-A 14 loci of MMR proficient strains when compared to wild-type strains, respectively. Overexpressing AtMLH1 in the AtPMS1-overproducing strain generated an increase in mutation rate comparable to that of AtPMS1 expression alone. Deletion of the C-terminal residues implicated in protein–protein interaction and including the putative endonuclease sequence of AtPMS1 completely eliminated the mutator phenotype. Taken together, these results indicate that the plant proteins affect yeast genomic stability, very possibly altering protein–protein interactions that are necessary to complete repair.

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Abbreviations

MMR:

Mismatch repair

At:

Arabidopsis thaliana

Sc:

Saccharomyces cerevisiae

Hs:

Homo sapiens

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Acknowledgments

We wish to thank Thomas Kunkel (National Institute of Environmental Health Sciences, Research Triangle Park, USA) for providing yeast strains used in recombinant expression experiments and mutation rate determination assays and Dmitry Gordenin (National Institute of Environmental Health Sciences, Research Triangle Park, USA) for providing YEp112SPGAL and YEp195SPGAL vectors. We would also like to include a special thanks to Rodrigo Gomez for his invaluable contribution to results discussion. This work was supported by grants from Agencia Nacional de Promoción Científica y Tecnológica (PICT 0458), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET; PIP 0018) and Universidad Nacional de Rosario (BIO 221). CPS is a member of the Researcher Career of CONICET. CG is a fellow of the same institution.

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  1. Centro de Estudios Fotosintéticos y Bioquímicos (CEFOBI), Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Suipacha 531, 2000, Rosario, Argentina

    Celina Galles & Claudia P. Spampinato

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  1. Celina Galles
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  2. Claudia P. Spampinato
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Correspondence to Claudia P. Spampinato.

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Galles, C., Spampinato, C.P. Yeast mutator phenotype enforced by Arabidopsis PMS1 expression. Mol Biol Rep 40, 2107–2114 (2013). https://doi.org/10.1007/s11033-012-2269-5

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