Transformation of <i>Schizosaccharomyces japonicus</i>

Aoki, Keita; Niki, Hironori

doi:10.1101/pdb.prot091850

Cited by 13 publications

(12 citation statements)

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“…The deletion of elp4 in S. japonicus was performed by electroporation using a polymerase chain reaction (PCR) product flanked by long homology arms generated by a three-step PCR as previously described ( 61 , 62 ). Mutageneses were performed using the Q5 Site-Directed Mutagenesis Kit (New England BioLabs).…”

Section: Methodsmentioning

confidence: 99%

Reciprocal regulation of TORC signaling and tRNA modifications by Elongator enforces nutrient-dependent cell fate

et al. 2019

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Nutrient availability has a profound impact on cell fate. Upon nitrogen starvation, wild-type fission yeast cells uncouple cell growth from cell division to generate small, round-shaped cells that are competent for sexual differentiation. The TORC1 (TOR complex 1) and TORC2 complexes exert opposite controls on cell growth and cell differentiation, but little is known about how their activity is coordinated. We show that transfer RNA (tRNA) modifications by Elongator are critical for this regulation by promoting the translation of both key components of TORC2 and repressors of TORC1. We further identified the TORC2 pathway as an activator of Elongator by down-regulating a Gsk3 (glycogen synthase kinase 3)–dependent inhibitory phosphorylation of Elongator. Therefore, a feedback control is operating between TOR complex (TORC) signaling and tRNA modification by Elongator to enforce the advancement of mitosis that precedes cell differentiation.

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Section: Methodsmentioning

confidence: 99%

Reciprocal regulation of TORC signaling and tRNA modifications by Elongator enforces nutrient-dependent cell fate

et al. 2019

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“…A 3-kb fragment was amplified from pTS7 and purified by using the QIAquick PCR Purification Kit (Qiagen, Tokyo, Japan). The DNA fragment was transformed in a heterothallic M-strain, NIG5872, by electroporation as described previously [ 18 ], and transformants were selected on YEA containing G418. Primers oTS9/oTS38 were used to confirm that the ORF of sxa2 + had been deleted.…”

Section: Methodsmentioning

confidence: 99%

“…In recent years, genetic methods for studying S. japonicus (e.g., transformation [ 18 , 19 ] and spore dissection [ 20 ]) and bioresources (e.g., heterothallic strains [ 15 ]) have become available. In addition, whole-genome sequences of S. japonicus were published by the Broad Institute in 2011 [ 21 ].…”

Section: Introductionmentioning

confidence: 99%

Elevated Sporulation Efficiency in Fission Yeast Schizosaccharomyces japonicus Strains Isolated from Drosophila

Seike

Sakata

Mizutani

et al. 2021

JoF

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The fission yeast Schizosaccharomyces japonicus, comprising S. japonicus var. japonicus and S. japonicus var. versatilis varieties, has unique characteristics such as striking hyphal growth not seen in other Schizosaccharomyces species; however, information on its diversity and evolution, in particular mating and sporulation, remains limited. Here we compared the growth and mating phenotypes of 17 wild strains of S. japonicus, including eight S. japonicus var. japonicus strains newly isolated from an insect (Drosophila). Unlike existing wild strains isolated from fruits/plants, the strains isolated from Drosophila sporulated at high frequency even under nitrogen-abundant conditions. In addition, one of the strains from Drosophila was stained by iodine vapor, although the type strain of S. japonicus var. japonicus is not stained. Sequence analysis further showed that the nucleotide and amino acid sequences of pheromone-related genes have diversified among the eight strains from Drosophila, suggesting crossing between S. japonicus cells of different genetic backgrounds occurs frequently in this insect. Much of yeast ecology remains unclear, but our findings suggest that insects such as Drosophila might be a good niche for mating and sporulation, and will provide a basis for the understanding of sporulation mechanisms via signal transduction, as well as the ecology and evolution of yeast.

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“…The general genetic methods used in this study were described previously (Aoki & Niki, 2017;Forsburg, 2003;Moreno et al, 1991).…”

Section: Strains and Mediamentioning

confidence: 99%

Yesprit and Yeaseq: Applications for designing primers and browsing sequences for research using the fourSchizosaccharomycesspecies

Wang

et al. 2021

Yeast

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The polymerase chain reaction (PCR)-based gene targeting method, which can delete a specific gene or introduce tags, has been widely utilized to study gene function in fission yeast. One of the critical steps in this method is to design primers for amplifying DNA fragments of deletion or tagging modules and for checking the integration of those DNA fragments at designated loci. Although the primer design tool Pombe PCR Primer Program (PPPP) is available for Schizosaccharomyces pombe, there is no such publicly available application for the other three fission yeast species, S. cryophilus, S. japonicus, and S. octosporus. Likewise, no application enabling DNA/protein sequence retrieval for these three fission yeast species is available either. Therefore, access to such functionality would substantially assist in retrieval of gene sequences of interest and primer design in these fission yeast species. In this report, we describe two applications for fission yeast study: Yesprit and Yeaseq.Yesprit is a primer design tool for strain construction using the PCR-based method, and Yeaseq is a sequence viewer that can acquire the DNA/protein sequences of specific genes. Both tools can be run on the Windows, macOS, and Linux platforms.We believe that the Yesprit and Yeaseq will facilitate research using the four fission yeast species. Take Aways• Yesprit designs primers for gene deletion/tagging in the four fission yeasts.• Gene deletion and tagging using Yesprit-designed primers verified its utility.• Yeaseq can retrieve DNA/protein sequences of the four fission yeast species.• Yeaseq helps construct temperature-sensitive mutants of S. japonicus. K E Y W O R D S fission yeast, PCR-based gene targeting, primer design tool, sequence viewer 1 | INTRODUCTION Fission yeasts are model organisms that have been extensively used in biological research because, although unicellular, they share many biological processes with metazoans, such as the eukaryotic cell cycle,

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Transformation of Schizosaccharomyces japonicus

Cited by 13 publications

References 1 publication

Reciprocal regulation of TORC signaling and tRNA modifications by Elongator enforces nutrient-dependent cell fate

Reciprocal regulation of TORC signaling and tRNA modifications by Elongator enforces nutrient-dependent cell fate

Elevated Sporulation Efficiency in Fission Yeast Schizosaccharomyces japonicus Strains Isolated from Drosophila

Yesprit and Yeaseq: Applications for designing primers and browsing sequences for research using the fourSchizosaccharomycesspecies

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