Over-expression and characterization of recombinant prefoldin from hyperthermophilic archaeum Pyrococcus furiosus in E. coli

Chen, Hui; Yang, Li-da; Zhang, Yi; Yang, Shengli

doi:10.1007/s10529-009-0156-5

Cited by 8 publications

(5 citation statements)

References 19 publications

(17 reference statements)

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“…For instance, PFDN expression protects against organic solvents toxicity by sequestrating unfolded proteins and thus cooperating with the bacterial chaperone systems to reduce the protein stress caused by exposure to these types of solvents ( Okochi et al., 2008 ). It also increases the tolerance to hyperthermia in these organisms, consistent with the role of archaeal PFDN in preventing protein aggregation and favoring a proper protein folding ( Chen et al., 2010 ) ( Figure 2 ). PFDN, unlike other archaeal chaperones, also rescues the cell growth inhibition caused by antibiotic (aminoglycoside) toxicity, probably through a decrease in the protein folding aggregation caused by this agent ( Peng et al., 2017 ).…”

Section: Introductionsupporting

confidence: 75%

A comprehensive analysis of prefoldins and their implication in cancer

2021

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Summary Prefoldins (PFDNs) are evolutionary conserved co-chaperones, initially discovered in archaea but universally present in eukaryotes. PFDNs are prevalently organized into hetero-hexameric complexes. Although they have been overlooked since their discovery and their functions remain elusive, several reports indicate they act as co-chaperones escorting misfolded or non-native proteins to group II chaperonins. Unlike the eukaryotic PFDNs which interact with cytoskeletal components, the archaeal PFDNs can bind and stabilize a wide range of substrates, possibly due to their great structural diversity. The discovery of the unconventional RPB5 interactor (URI) PFDN-like complex (UPC) suggests that PFDNs have versatile functions and are required for different cellular processes, including an important role in cancer. Here, we summarize their functions across different species. Moreover, a comprehensive analysis of PFDNs genomic alterations across cancer types by using large-scale cancer genomic data indicates that PFDNs are a new class of non-mutated proteins significantly overexpressed in some cancer types.

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Section: Introductionsupporting

confidence: 75%

A comprehensive analysis of prefoldins and their implication in cancer

2021

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“…It may because of the protection from HSPs is only short-acting [12] that once the duration of thermal stress is enhanced, the cells ultimately undergo cell death. Although the overexpression of HSP has been applied to improve the thermotolerance [49,50], the effects are not ideal. We assumed that preventing protein misfolding by OSR is more important than the repair by HSR after damage.…”

Section: Discussionmentioning

confidence: 99%

Augmented peroxisomal ROS buffering capacity renders oxidative and thermal stress cross-tolerance in yeast

Lin

et al. 2021

Microb Cell Fact

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Background Thermotolerant yeast has outstanding potential in industrial applications. Komagataella phaffii (Pichia pastoris) is a common cell factory for industrial production of heterologous proteins. Results Herein, we obtained a thermotolerant K. phaffii mutant G14 by mutagenesis and adaptive evolution. G14 exhibited oxidative and thermal stress cross-tolerance and high heterologous protein production efficiency. The reactive oxygen species (ROS) level and lipid peroxidation in G14 were reduced compared to the parent. Oxidative stress response (OSR) and heat shock response (HSR) are two major responses to thermal stress, but the activation of them was different in G14 and its parent. Compared with the parent, G14 acquired the better performance owing to its stronger OSR. Peroxisomes, as the main cellular site for cellular ROS generation and detoxification, had larger volume in G14 than the parent. And, the peroxisomal catalase activity and expression level in G14 was also higher than that of the parent. Excitingly, the gene knockdown of CAT encoding peroxisomal catalase by dCas9 severely reduced the oxidative and thermal stress cross-tolerance of G14. These results suggested that the augmented OSR was responsible for the oxidative and thermal stress cross-tolerance of G14. Nevertheless, OSR was not strong enough to protect the parent from thermal stress, even when HSR was initiated. Therefore, the parent cannot recover, thereby inducing the autophagy pathway and resulting in severe cell death. Conclusions Our findings indicate the importance of peroxisome and the significance of redox balance in thermotolerance of yeasts.

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“…Genes encoding prefoldin, chaperonin (HSP60), sHSP, and extracellular α-amylase from P. furiosus (PFA) were PCRamplified from plasmids constructed in previous studies (Chen et al 2006(Chen et al , 2010. Primers used to amplify these g e n e s w e r e a s f o l l o w s : p r e f o l d i n : s e n s e , 5 ′ -GCCCATGGAAAACAATAAGGAAT-3′ (NcoI underlined); antisense, 5′-ACGCGTCGACGGAGCTCGAATTCTCATC-3′ (SalI underlined); HSP60: sense, 5′-ACTATCATATGGC CCAGTTAGCAGGC-3′ (NdeI underlined); antisense, 5′-CAGCTCGAGCTTGCTGGCAGCGATGAC-3′ (XhoI underlined); sHSP: sense, 5′-GCCCATGGTTCGTCGTATC-3′ (NcoI underlined); antisense, 5′ -AACG TC GAC TATTCAACTTT AACTTC-3′ (SalI underlined).…”

Section: Constructions Of Recombinant Plasmidsmentioning

confidence: 99%

“…Previous studies indicated that archaeal prefoldins were able to protect the aggregation of diverse heterogeneous proteins (Iizuka et al 2008;Leroux et al 1999;Laksanalamai et al 2006;Okochi et al 2002;Lundin et al 2004;Hongo et al 2012). Furthermore, overexpression of prefoldin from hyperthermophilic archaea was able to efficiently increase tolerance of E. coli hosts to different environmental stresses, such as the presence of an organic solvent (Okochi et al 2008) or high temperature (Chen et al 2010).…”

Section: Introductionmentioning

confidence: 99%

Co-expression of chaperones from P. furiosus enhanced the soluble expression of the recombinant hyperthermophilic α-amylase in E. coli

Peng

Chu

et al. 2016

Cell Stress and Chaperones

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The extracellular α-amylase from the hyperthermophilic archaeum Pyrococcus furiosus (PFA) is extremely thermostable and of an industrial importance and interest. PFA aggregates and accumulates as insoluble inclusion bodies when expressed as a heterologous protein at a high level in Escherichia coli. In the present study, we investigated the roles of chaperones from P. furiosus in the soluble expression of recombinant PFA in E. coli. The results indicate that coexpression of PFA with the molecular chaperone prefoldin alone significantly increased the soluble expression of PFA. Although, co-expression of other main chaperone components from P. furiosus, such as the small heat shock protein (sHSP) or chaperonin (HSP60), was also able to improve the soluble expression of PFA to a certain extent. Co-expression of chaperonin or sHSP in addition to prefoldin did not further increase the soluble expression of PFA. This finding emphasizes the biotechnological potentials of the molecular chaperone prefoldin from P. furiosus, which may facilitate the production of recombinant PFA.

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Over-expression and characterization of recombinant prefoldin from hyperthermophilic archaeum Pyrococcus furiosus in E. coli

Cited by 8 publications

References 19 publications

A comprehensive analysis of prefoldins and their implication in cancer

A comprehensive analysis of prefoldins and their implication in cancer

Augmented peroxisomal ROS buffering capacity renders oxidative and thermal stress cross-tolerance in yeast

Co-expression of chaperones from P. furiosus enhanced the soluble expression of the recombinant hyperthermophilic α-amylase in E. coli

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