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Giant yeast cells with nonrecyclable ribonucleotide reductase.
MedLine Citation:
PMID:  21442328     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Ribonucleotide reductase (RNR) catalyzes the reduction of ribonucleotides to deoxyribonucleotides and thereby provides the precursors required for DNA synthesis and repair. In an attempt to test cell resistance to a permanent replicational stress, we constructed a mutant Saccharomyces cerevisiae strain containing exclusively nonrecyclable catalytic subunits of RNR that become inactivated following the reduction of one ribonucleoside diphosphate. In this rnr1C883A rnr3Δ mutant, the synthesis of each deoxyribonucleotide thus requires the production of one Rnr1C883A protein, which means that 26 million Rnr1C883A proteins (half the protein complement of a wild-type cell) have to be produced during each cell cycle. rnr1C883A rnr3Δ cells grow under constant replicational stress, as evidenced by the constitutive activation of the checkpoint effector Rad53, and their S phase is considerably extended compared to the wild type. rnr1C883A rnr3Δ mutants also display additional abnormalities such as a median cell volume increased by a factor of 8, and the presence of massive inclusion bodies. However, they exhibit a good plating efficiency and can be propagated indefinitely. rnr1C883A rnr3Δ cells, which can be used as a protein overexpression system, thus illustrate the robustness of S. cerevisiae to multiple physiological parameters.
Authors:
Emilie Ma; Arach Goldar; Jean-Marc Verbavatz; Marie-Claude Marsolier-Kergoat
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-3-27
Journal Detail:
Title:  Molecular genetics and genomics : MGG     Volume:  -     ISSN:  1617-4623     ISO Abbreviation:  -     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-3-28     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101093320     Medline TA:  Mol Genet Genomics     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Institut de Biologie et de Technologies de Saclay, Service de Biologie Intégrative et de Génétique Moléculaire, Bât. 144, CEA/Saclay, 91191, Gif-sur-Yvette Cedex, France.
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