Pim1, a MAP kinase involved in cell wall integrity in Pichia pastoris

Ic, Cosano; Martı́n, Humberto; Flández, Marta; Nombela, César; Molina, Marı́a

doi:10.1007/s004380100452

Cited by 12 publications

(4 citation statements)

References 42 publications

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“…For example, the disruption of ARG1, ARG2, ARG3, HIS1, HIS2, HIS5, and HIS6 in P. pastoris GS115 strain with flanking arms that range from 200 bp to 900 bp is considerably efficient at frequencies of 44% to 90% [7]. However, the deletion of PIM [8] and OCH1 [9] from the same strain that contains both flanking arms that were longer than 1 kb occurred at a frequency of <1%. To date, this locus specific phenomenon is not well understood.…”

Section: Introductionmentioning

confidence: 99%

Enhancement of the Gene Targeting Efficiency of Non-Conventional Yeasts by Increasing Genetic Redundancy

Chen

Sun

et al. 2013

PLoS ONE

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In contrast to model yeasts, gene targeting efficiencies of non-conventional yeasts are usually low, which greatly limits the research and applications of these organisms. In this study, we aimed to enhance the gene targeting efficiency of non-conventional yeasts by improving the fitness of mutant strains, particularly by increasing the genetic redundancy of host cells. To demonstrate this process, OCH1 gene deletion in Pichia pastoris was performed. Extra copies of the OCH1 gene on a helper plasmid were provided for the P. pastoris GS115 strain before the native OCH1 gene in the genomic DNA was knocked out. The redundancy in OCH1 gene significantly eliminated the growth defects of the och1 mutant and increased the deletion efficiency of the OCH1 gene by two orders of magnitude with the same length of homologous flanks. The same strategy was used to delete the KU70 and SGS1 genes. The targeting efficiencies of KU70 and SGS1 were increased by 1- and 23-fold, respectively. Therefore, this study provided an efficient strategy for the deletion of “stubborn” genes in non-conventional yeasts. This study further showed that cellular fitness is potentially an important factor that can limit the efficiency of gene targeting.

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Section: Introductionmentioning

confidence: 99%

Enhancement of the Gene Targeting Efficiency of Non-Conventional Yeasts by Increasing Genetic Redundancy

Chen

Sun

et al. 2013

PLoS ONE

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“…So far, several homologous proteins of the CWI pathway have been identified in K. phaffii. In 2001, the K. phaffii homolog of Slt2 was characterized and named Pim1 (P. pastoris cell integrity MAPK) (Cosano et al, 2001). Similar to what has been published for ScSlt2, Pim1/Slt2 is dually phosphorylated, and thereby activated in response to heat stress, caffeine and agents that affect the integrity of the fungal cell wall.…”

Section: The Cell Wallmentioning

confidence: 82%

Komagataella phaffii as Emerging Model Organism in Fundamental Research

et al. 2021

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Komagataella phaffii (Pichia pastoris) is one of the most extensively applied yeast species in pharmaceutical and biotechnological industries, and, therefore, also called the biotech yeast. However, thanks to more advanced strain engineering techniques, it recently started to gain attention as model organism in fundamental research. So far, the most studied model yeast is its distant cousin, Saccharomyces cerevisiae. While these data are of great importance, they limit our knowledge to one organism only. Since the divergence of the two species 250 million years ago, K. phaffii appears to have evolved less rapidly than S. cerevisiae, which is why it remains more characteristic of the common ancient yeast ancestors and shares more features with metazoan cells. This makes K. phaffii a valuable model organism for research on eukaryotic molecular cell biology, a potential we are only beginning to fully exploit. As methylotrophic yeast, K. phaffii has the intriguing property of being able to efficiently assimilate methanol as a sole source of carbon and energy. Therefore, major efforts have been made using K. phaffii as model organism to study methanol assimilation, peroxisome biogenesis and pexophagy. Other research topics covered in this review range from yeast genetics including mating and sporulation behavior to other cellular processes such as protein secretion, lipid biosynthesis and cell wall biogenesis. In this review article, we compare data obtained from K. phaffii with S. cerevisiae and other yeasts whenever relevant, elucidate major differences, and, most importantly, highlight the big potential of using K. phaffii in fundamental research.

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“…Given that Calcofluor white, SDS, vanadate and caffeine also induce phosphorylation and activation of Slt2 (54,57,58), it is intriguing that they do not affect rDNA silencing. Calcofluor white is a chitin antagonist and has a much weaker binding affinity toward β-1,3-glucan than Congo red (59).…”

Section: Discussionmentioning

confidence: 99%

The β-1,3-glucanosyltransferase Gas1 regulates Sir2-mediated rDNA stability inSaccharomyces cerevisiae

Kim

Chang

et al. 2014

Nucleic Acids Res

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In Saccharomyces cerevisiae, the stability of highly repetitive rDNA array is maintained through transcriptional silencing. Recently, a β-1,3-glucanosyltransferase Gas1 has been shown to play a significant role in the regulation of transcriptional silencing in S. cerevisiae. Here, we show that the gas1Δ mutation increases rDNA silencing in a Sir2-dependent manner. Remarkably, the gas1Δ mutation induces nuclear localization of Msn2/4 and stimulates the expression of PNC1, a gene encoding a nicotinamidase that functions as a Sir2 activator. The lack of enzymatic activity of Gas1 or treatment with a cell wall-damaging agent, Congo red, exhibits effects similar to those of the gas1Δ mutation. Furthermore, the loss of Gas1 or Congo red treatment lowers the cAMP-dependent protein kinase (PKA) activity in a cell wall integrity MAP kinase Slt2-dependent manner. Collectively, our results suggest that the dysfunction of Gas1 plays a positive role in the maintenance of rDNA integrity by decreasing PKA activity and inducing the accumulation of Msn2/4 in the nucleus. It seems that nuclear-localized Msn2/4 stimulate the expression of Pnc1, thereby enhancing the association of Sir2 with rDNA and promoting rDNA stability.

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Pim1, a MAP kinase involved in cell wall integrity in Pichia pastoris

Cited by 12 publications

References 42 publications

Enhancement of the Gene Targeting Efficiency of Non-Conventional Yeasts by Increasing Genetic Redundancy

Enhancement of the Gene Targeting Efficiency of Non-Conventional Yeasts by Increasing Genetic Redundancy

Komagataella phaffii as Emerging Model Organism in Fundamental Research

The β-1,3-glucanosyltransferase Gas1 regulates Sir2-mediated rDNA stability inSaccharomyces cerevisiae

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