title: Polypeptone induces dramatic cell lysis in ura4 deletion mutants of fission yeast.
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.
authors.affiliation: Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Matsue, Japan.
authors.forename: Yuzy
authors.initials: Y
authors.lastname: Matsuo
authors.forename: Kouhei
authors.initials: K
authors.lastname: Nishino
authors.forename: Kouhei
authors.initials: K
authors.lastname: Mizuno
authors.forename: Takashi
authors.initials: T
authors.lastname: Akihiro
authors.forename: Takashi
authors.initials: T
authors.lastname: Toda
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Matsuo
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Kaino
authors.forename: Makoto
authors.initials: M
authors.lastname: Kawamukai
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism; D002853:Chromatography, Liquid; D017353:Gene Deletion; D005819:Genetic Markers; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism; D013058:Mass Spectrometry; D008957:Models, Genetic; D009995:Osmosis; D010461:Peptones / Q000494:pharmacology*; D010641:Phenotype; D012441:Saccharomyces cerevisiae; D012568:Schizosaccharomyces / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D005819:Genetic Markers; D008164:Luminescent Proteins; D010461:Peptones; D029702:Schizosaccharomyces pombe Proteins; C400810:ura4 protein, S pombe; C054562:yellow fluorescent protein, Bacteria; D000469:Alkaline Phosphatase; D004693:Glucan Endo-1,3-beta-D-Glucosidase
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