A mutated dph3 gene causes sensitivity of Schizosaccharomyces pombe cells to cytotoxic agents

Autor: Desirée Villahermosa, Karen Knapp, Oliver Fleck
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
Genetic Markers
Saccharomyces cerevisiae Proteins
Mutant
Saccharomyces cerevisiae
Codon
Initiator
DNA damage response
03 medical and health sciences
chemistry.chemical_compound
Eukaryotic translation
Peptide Elongation Factor 2
Genome stability
Gene Expression Regulation
Fungal
Schizosaccharomyces
Genetics
Hydroxyurea
Histidine
Gene
biology
Cytotoxins
Diphthamide
Wild type
Elongator
General Medicine
biology.organism_classification
Methyl Methanesulfonate
Molecular biology
Translation elongation factor eEF2
Methyl methanesulfonate
Sordarin
Transcriptional interference
030104 developmental biology
chemistry
Indenes
Genetic Loci
Schizosaccharomyces pombe
tRNA modifications
MutS Homolog 3 Protein
Mutation
Original Article
Schizosaccharomyces pombe Proteins
Genetic Engineering
Zdroj: Current Genetics
ISSN: 0172-8083
Popis: Dph3 is involved in diphthamide modification of the eukaryotic translation elongation factor eEF2 and in Elongator-mediated modifications of tRNAs, where a 5-methoxycarbonyl-methyl moiety is added to wobble uridines. Lack of such modifications affects protein synthesis due to inaccurate translation of mRNAs at ribosomes. We have discovered that integration of markers at the msh3 locus of Schizosaccharomyces pombe impaired the function of the nearby located dph3 gene. Such integrations rendered cells sensitive to the cytotoxic drugs hydroxyurea and methyl methanesulfonate. We constructed dph3 and msh3 strains with mutated ATG start codons (ATGmut), which allowed investigating drug sensitivity without potential interference by marker insertions. The dph3-ATGmut and a dph3::loxP-ura4-loxM gene disruption strain, but not msh3-ATGmut, turned out to be sensitive to hydroxyurea and methyl methanesulfonate, likewise the strains with cassettes integrated at the msh3 locus. The fungicide sordarin, which inhibits diphthamide modified eEF2 of Saccharomyces cerevisiae, barely affected survival of wild type and msh3Δ S. pombe cells, while the dph3Δ mutant was sensitive. The msh3-ATG mutation, but not dph3Δ or the dph3-ATG mutation caused a defect in mating-type switching, indicating that the ura4 marker at the dph3 locus did not interfere with Msh3 function. We conclude that Dph3 is required for cellular resistance to the fungicide sordarin and to the cytotoxic drugs hydroxyurea and methyl methanesulfonate. This is likely mediated by efficient translation of proteins in response to DNA damage and replication stress. Electronic supplementary material The online version of this article (doi:10.1007/s00294-017-0711-x) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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