RRC ID 21264
Author Matsuo Y, Nishino K, Mizuno K, Akihiro T, Toda T, Matsuo Y, Kaino T, Kawamukai M.
Title Polypeptone induces dramatic cell lysis in ura4 deletion mutants of fission yeast.
Journal PLoS ONE
Abstract Polypeptone is widely excluded from Schizosaccharomyces pombe growth medium. However, the reasons why polypeptone should be avoided have not been documented. Polypeptone dramatically induced cell lysis in the ura4 deletion mutant when cells approached the stationary growth phase, and this phenotype was suppressed by supplementation of uracil. To determine the specificity of this cell lysis phenotype, we created deletion mutants of other genes involved in de novo biosynthesis of uridine monophosphate (ura1, ura2, ura3, and ura5). Cell lysis was not observed in these gene deletion mutants. In addition, concomitant disruption of ura1, ura2, ura3, or ura5 in the ura4 deletion mutant suppressed cell lysis, indicating that cell lysis induced by polypeptone is specific to the ura4 deletion mutant. Furthermore, cell lysis was also suppressed when the gene involved in coenzyme Q biosynthesis was deleted. This is likely because Ura3 requires coenzyme Q for its activity. The ura4 deletion mutant was sensitive to zymolyase, which mainly degrades (1,3)-beta-D glucan, when grown in the presence of polypeptone, and cell lysis was suppressed by the osmotic stabiliser, sorbitol. Finally, the induction of cell lysis in the ura4 deletion mutant was due to the accumulation of orotidine-5-monophosphate. Cell wall integrity was dramatically impaired in the ura4 deletion mutant when grown in the presence of polypeptone. Because ura4 is widely used as a selection marker in S. pombe, caution needs to be taken when evaluating phenotypes of ura4 mutants.
Volume 8(3)
Pages e59887
Published 2013
DOI 10.1371/journal.pone.0059887
PII PONE-D-12-34903
PMID 23555823
PMC PMC3605382
MeSH Alkaline Phosphatase / metabolism Bacterial Proteins / metabolism Cell Wall / metabolism Chromatography, Liquid Gene Deletion* Genetic Markers Glucan Endo-1,3-beta-D-Glucosidase / metabolism Luminescent Proteins / metabolism Mass Spectrometry Models, Genetic Osmosis Peptones / pharmacology* Phenotype Saccharomyces cerevisiae Schizosaccharomyces / genetics* Schizosaccharomyces pombe Proteins / genetics*
IF 2.766
Times Cited 11
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Yeast FY7520(L968)