The <i>Yarrowia lipolytica PAH1</i> homologue contributes but is not required for triacylglycerol biosynthesis during growth on glucose

Ukey, Rahul; Carmon, Taylor; Hardman, Derell; Hill, Na'Taja; Fakas, Stylianos

doi:10.1002/yea.3447

Cited by 5 publications

(5 citation statements)

References 34 publications

(84 reference statements)

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“…Decreased levels of DAG and TAG were observed in the Mopah1 mutant, which has also been observed in pah1 mutants of S. cerevisiae, F. graminearum, and Yarrowia lipolytica (Karanasios et al, 2010;Oh et al, 2017;Ukey et al, 2020). The reduction of TAG levels leads to a reduced level of lipid droplets in pah1 mutants of Tetrahymena thermophila and F. graminearum (Liu, Yun, et al, 2019a;Liu, Liang, et al,2019b;Pillai et al, 2017;Shi et al, 2018).…”

Section: Discussionmentioning

confidence: 75%

The MoPah1 phosphatidate phosphatase is involved in lipid metabolism, development, and pathogenesis in Magnaporthe oryzae

Zhao

Sun

et al. 2022

Molecular Plant Pathology

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As with the majority of the hemibiotrophic fungal pathogens, the rice blast fungus Magnaporthe oryzae uses highly specialized infection structures called appressoria for plant penetration. Appressoria differentiated from germ tubes rely on enormous turgor pressure to directly penetrate the plant cell, in which process lipid metabolism plays a critical role. In this study, we characterized the MoPAH1 gene in M. oryzae, encoding a putative highly conserved phosphatidate phosphatase. The expression of MoPAH1 was up-regulated during plant infection. The MoPah1 protein is expressed at all developmental and infection stages, and is localized to the cytoplasm. Disruption of MoPAH1 causes pleiotropic defects in vegetative growth, sporulation, and heat tolerance. The lipid profile is significantly altered in the Mopah1 mutant. Lipidomics assays showed that the level of phosphatidic acid (PA) was increased in the mutant, which had reduced levels of diacylglycerol and triacylglycerol. Using a PA biosensor, we showed that the increased level of PA in the Mopah1 mutant was primarily accumulated in the vacuole. The Mopah1 mutant was blocked in both conidiation and the formation of appressorium-like structures at hyphal tips. It was nonpathogenic and failed to cause any blast lesions on rice and barley seedlings. RNA sequencing analysis revealed that MoPah1 regulates the expression of transcription factors critical for various developmental and infection-related processes. The Mopah1 mutant was reduced in the expression and phosphorylation of Pmk1 MAP kinase and delayed in autophagy. Our study demonstrates that MoPah1 is necessary for lipid metabolism, fungal development, and pathogenicity in M. oryzae.

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Section: Discussionmentioning

confidence: 75%

The MoPah1 phosphatidate phosphatase is involved in lipid metabolism, development, and pathogenesis in Magnaporthe oryzae

Zhao

Sun

et al. 2022

Molecular Plant Pathology

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“…[48,49]. It has been elucidated that PAH1 controls the biosynthesis of TAG and phospholipid membranes and the abundance of lipid signaling molecules [50]. Similarly, PAH1 in the yeast catalyzes the Mg 2+ dependent dephosphorylation of PA, producing DAG at the nuclear/ER membrane.…”

Section: Discussionmentioning

confidence: 99%

Tracing Key Molecular Regulators of Lipid Biosynthesis in Tuber Development of Cyperus esculentus Using Transcriptomics and Lipidomics Profiling

Wang

Jing

Ahmad

et al. 2021

Genes

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Cyperus esculentus is widely representing one of the important oil crops around the world, which provides valuable resources of edible tubers called tiger nut. The chemical composition and high ability to produce fats emphasize the role of tiger nut in promoting oil crop productivity. However, the underlying molecular mechanism of the production and accumulation of lipids in tiger nut development still remains unclear. Here, we conducted comprehensive transcriptomics and lipidomics analyses at different developmental stages of tuber in Cyperus esculentus. Lipidomic analyses confirmed that the accumulation of lipids including glycolipids, phospholipids, and glycerides were significantly enriched during tuber development from early to mature stage. The proportion of phosphatidylcholines (PC) declined during all stages and phosphatidyl ethanolamine (PE) was significantly declined in early and middle stages. These findings implied that PC is actively involved in triacylglycerol (TAG) biosynthesis during the tubers development, whereas PE may participate in TAG metabolism during early and middle stages. Comparative transcriptomics analyses indicated several genomic and metabolic pathways associated with lipid metabolism during tuber development in tiger nut. The Pearson correlation analysis showed that TAG synthesis in different developmental stages was attributed to 37 candidate transcripts including CePAH1. The up-regulation of diacylglycerol (DAG) and oil content in yeast, resulted from the inducible expression of exogenous CePAH1 confirmed the central role of this candidate gene in lipid metabolism. Our results demonstrated the foundation of an integrative metabolic model for understanding the molecular mechanism of tuber development in tiger nut, in which lipid biosynthesis plays a central role.

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“…According to Guerfal et al [24], knockout of PAH1 will disrupt the TAG synthesis pathway and direct the fatty acid flux to phospholipids, leading to ER membrane proliferation. This ER expansion is expected to enhance the membrane protein accumulation levels [24][25][26]33]. We hypothesized that an expanded ER would increase the processing efficiency of the secreted proteins and thus improve the secretion of T4 lysozyme.…”

Section: Co-expression Of Scste13 and Scerv29 In Pah1 Strain Improve ...mentioning

confidence: 99%

“…In the secretory pathway, the ER and Golgi play a key role in protein folding, post-translation modification, and quality control of translated protein. Deleting the phosphatidic acid phosphatase gene, PAH1 , impaired triacylglycerol formation and sterol ester storage, and triggered proliferation of the ER membrane in Y. lipolytica [ 24 , 25 ]. The same strategy was successfully used to enhance protein secretion in S. cerevisiae [ 26 ].…”

Section: Introductionmentioning

confidence: 99%

Engineering heterologous enzyme secretion in Yarrowia lipolytica

Wang

Blenner

2022

Microb Cell Fact

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Background Eukaryotic cells are often preferred for the production of complex enzymes and biopharmaceuticals due to their ability to form post-translational modifications and inherent quality control system within the endoplasmic reticulum (ER). A non-conventional yeast species, Yarrowia lipolytica, has attracted attention due to its high protein secretion capacity and advanced secretory pathway. Common means of improving protein secretion in Y. lipolytica include codon optimization, increased gene copy number, inducible expression, and secretory tag engineering. In this study, we develop effective strategies to enhance protein secretion using the model heterologous enzyme T4 lysozyme. Results By engineering the commonly used native lip2prepro secretion signal, we have successfully improved secreted T4 lysozyme titer by 17-fold. Similar improvements were measured for other heterologous proteins, including hrGFP and $$\alpha$$ α -amylase. In addition to secretion tag engineering, we engineered the secretory pathway by expanding the ER and co-expressing heterologous enzymes in the secretion tag processing pathway, resulting in combined 50-fold improvement in T4 lysozyme secretion. Conclusions Overall, our combined strategies not only proved effective in improving the protein production in Yarrowia lipolytica, but also hint the possible existence of a different mechanism of secretion regulation in ER and Golgi body in this non-conventional yeast.

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The Yarrowia lipolytica PAH1 homologue contributes but is not required for triacylglycerol biosynthesis during growth on glucose

Cited by 5 publications

References 34 publications

The MoPah1 phosphatidate phosphatase is involved in lipid metabolism, development, and pathogenesis in Magnaporthe oryzae

The MoPah1 phosphatidate phosphatase is involved in lipid metabolism, development, and pathogenesis in Magnaporthe oryzae

Tracing Key Molecular Regulators of Lipid Biosynthesis in Tuber Development of Cyperus esculentus Using Transcriptomics and Lipidomics Profiling

Engineering heterologous enzyme secretion in Yarrowia lipolytica

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