Survey of molecular chaperone requirement for the biosynthesis of hamster polyomavirus VP1 protein in Saccharomyces cerevisiae

Valaviciute, Monika; Norkienė, Milda; Goda, Karolis; Slibinskas, Rimantas; Gedvilaitė, Alma

doi:10.1007/s00705-016-2846-3

Cited by 3 publications

(2 citation statements)

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“…It is believed that chaperone action is crucial to mitigate negative impacts related to protein misfolding. Valaviciute et al (2016) evaluated the effect of overexpression and mild downregulation of HSP90, HSP70, and HSP40 chaperones and co‐chaperones during recombinant expression of VP1 hamster polyomavirus protein in yeast. Downregulation of cytosolic chaperones such as SSA1/SSA2, SSA3/SSA4, HSP82, and HSC82 had a negative effect on VP1‐EGFP levels.…”

Section: Discussionmentioning

confidence: 99%

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Proteome profiling and vector yield optimization in a recombinant adeno‐associated virus‐producing yeast model

et al. 2020

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Section: Discussionmentioning

confidence: 99%

“…misfolding Valaviciute et al (2016). evaluated the effect of overexpression and mild downregulation of HSP90, HSP70, and HSP40 chaperones and co-chaperones during recombinant expression of VP1 hamster polyomavirus protein in yeast.…”

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confidence: 99%

Proteome profiling and vector yield optimization in a recombinant adeno‐associated virus‐producing yeast model

et al. 2020

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Mutations of Kluyveromyces lactis dolichol kinase enhances secretion of recombinant proteins

Žiogienė

Valaviciute

Norkienė

et al. 2019

FEMS Yeast Research

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Although there are similarities in the core steps of the secretion pathway from yeast to higher eukaryotes, significant functional differences exist even among diverse yeast species. Here, we used next-generation sequencing to identify two mutations in the Kluyveromyces lactis KlSEC59 gene, encoding dolichol kinase (DK), which are responsible for an enhanced secretion phenotype in a previously isolated mutant, MD2/1-9. Compared with the temperature-sensitive Saccharomyces cerevisiae sec59-1 mutant, which exhibits reduced N-glycosylation and decreased secretory efficacy, the identified K. lactis DK mutations had fewer effects on glycosylation, as well as on survival at high temperature and cell wall integrity. Moreover, despite some glycosylation defects, double DK mutations (G405S and I419S) in the K. lactis mutant strain demonstrated three times the level of recombinant α-amylase secretion as the wild-type strain. Overexpression of potential suppressors KlMNN10, KlSEL1, KlERG20, KlSRT1, KlRER2, KlCAX4, KlLPP1 and KlDPP1 in the DK-mutant strain restored carboxypeptidase Y glycosylation to different extents and, with the exception of KISRT1, reduced α-amylase secretion to levels observed in wild-type cells. Our results suggest that enhanced secretion related to reduced activity of mutant DK in K. lactis results from mild glycosylation changes that affect activity of other proteins in the secretory pathway.

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Dolichol kinases from yeast, nematode and human can replace each other and exchange their domains creating active chimeric enzymes in yeast

Ziogiene,

Burdulis,

Timinskas

et al. 2024

PLoS ONE

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Protein glycosylation is a fundamental modification crucial for numerous intra- and extracellular functions in all eukaryotes. The phosphorylated dolichol (Dol-P) is utilized in N-linked protein glycosylation and other glycosylation pathways. Dolichol kinase (DK) plays a key role in catalyzing the phosphorylation of dolichol. The glycosylation patterns in the Kluyveromyces lactis DK mutant revealed that the yeast well tolerated a minor deficiency in Dol-P by adjusting protein glycosylation. Comparative analysis of sequences of DK homologs from different species of eukaryotes, archaea and bacteria and AlphaFold3 structural model studies, allowed us to predict that DK is most likely composed of two structural/functional domains. The activity of predicted K. lactis DK C-terminal domain expressed from the single copy in the chromosome was not sufficient to keep protein glycosylation level necessary for survival of K. lactis. However, the glycosylation level was partially restored by additionally provided and overexpressed N- or C-terminal domain. Moreover, co-expression of the individual N-and C-terminal domains restored the glycosylation of vacuolar carboxypeptidase Y in both K. lactis and Saccharomyces cerevisiae. Despite the differences in length and non-homologous sequences of the N-terminal domains the human and nematode Caenorhabditis elegans DKs successfully complemented DK functions in both yeast species. Additionally, the N-terminal domains of K. lactis and C. elegans DK could functionally substitute for one another, creating active chimeric enzymes. Our results suggest that while the C-terminal domain remains crucial for DK activity, the N-terminal domain may serve not only as a structural domain but also as a possible regulator of DK activity.

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Survey of molecular chaperone requirement for the biosynthesis of hamster polyomavirus VP1 protein in Saccharomyces cerevisiae

Cited by 3 publications

References 64 publications

Proteome profiling and vector yield optimization in a recombinant adeno‐associated virus‐producing yeast model

Proteome profiling and vector yield optimization in a recombinant adeno‐associated virus‐producing yeast model

Mutations of Kluyveromyces lactis dolichol kinase enhances secretion of recombinant proteins

Dolichol kinases from yeast, nematode and human can replace each other and exchange their domains creating active chimeric enzymes in yeast

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