Over-expression of Caj1, a plasma membrane associated J-domain protein in Saccharomyces cerevisiae, stabilizes amino acid permeases

Dobriyal, Neha; Sagarika, Preeti; Shrivastava, A.; Verma, Amit; Islam, Zeyaul; Gupta, Poonam; Mochizuki, Takahiro; Abe, Fumiyoshi; Sahi, Chandan

doi:10.1016/j.bbamem.2020.183435

Cited by 6 publications

(15 citation statements)

References 57 publications

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“…We also find that the interaction of Caj1 with PA lipids is pH dependent. These results are striking in the context of recent in vivo data, which suggests that Caj1 may be involved in membrane quality control 16, 17 . While it is not currently known how Caj1 performs its function, it has been reported that Caj1 is partially localized to the yeast plasma membrane 17 .…”

Section: Introductionsupporting

confidence: 55%

“…These results are striking in the context of recent in vivo data, which suggests that Caj1 may be involved in membrane quality control 16, 17 . While it is not currently known how Caj1 performs its function, it has been reported that Caj1 is partially localized to the yeast plasma membrane 17 . Our observation that Caj1 displays a high affinity for lipid bilayers containing phosphatidic acid provides insights into how this Hsp40 may be specifically targeted to the cell membrane.…”

Section: Introductionsupporting

confidence: 55%

“…Caj1 is Hsp40 heat shock protein belonging to the E. coli DnaJ family. While its exact biological role in the cell context remains unclear, recent in vivo work has suggested that Caj1 may be involved in membrane protein quality control by an as-yet uncharacterized mechanism 16, 17, 26 . Besides Caj1, we also identify the phosphatidylinositol (PI)-interacting proteins Pdr16, Pik1 and Inp53.…”

Section: Resultsmentioning

confidence: 99%

“…The biological role of Caj1 in yeast remains unclear, although recent work has shown that it may be involved in membrane protein quality control 16,17 . Importantly, microscopy studies on S. cerevisiae expressing endogenous levels of GFP-tagged Caj1 also showed that Caj1 localizes to the plasma membrane 17 . Our current study adds a novel dimension to this previous finding, as we show in vitro that the recruitment of Caj1 to membranes requires phosphatidic acid.…”

Section: What Is the Function Of Caj1 In Vivo?mentioning

confidence: 99%

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Nanodiscs-based proteomics identify Caj1 as an Hsp40 with affinity for phosphatidic acid lipids

Zhang

Young

Foster

et al. 2021

Preprint

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Many soluble proteins interact with membranes to perform important biological functions, including signal transduction, regulation, transport, trafficking and biogenesis. Despite their importance, these protein-membrane interactions are difficult to characterize due to their often-transient nature as well as phospholipids' poor solubility in aqueous solution. Here, we employ nanodiscs - small, water-soluble patches of lipid bilayer encircled with amphipathic scaffold proteins - along with quantitative proteomics to identify lipid-binding proteins in S. cerevisiae. Using nanodiscs reconstituted with yeast total lipid extracts or only phosphatidylethanolamine (PE-nanodiscs), we capture several known membrane-interacting proteins, including the Rab GTPases Sec4 and Ypt1, which play key roles in vesicle trafficking. Utilizing PE-nanodiscs enriched with phosphatidic acid (PEPA-nanodiscs), we specifically capture a member of the Hsp40/J-protein family, Caj1, whose function has recently been linked to membrane protein quality control. We show that Caj1 interaction with liposomes containing PA is modulated by pH and PE lipids, and depends on two patches of positively charged residues near the C-terminus of the protein. The protein Caj1 is the first example of an Hsp40/J-domain protein with affinity for membranes and phosphatidic acid lipid specificity. These findings highlight the utility of the nanodisc system to identify and characterize protein-lipid interactions that may not be evident using other methods.

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Section: Introductionsupporting

confidence: 55%

Section: Introductionsupporting

confidence: 55%

Section: Resultsmentioning

confidence: 99%

Section: What Is the Function Of Caj1 In Vivo?mentioning

confidence: 99%

See 2 more Smart Citations

Nanodiscs-based proteomics identify Caj1 as an Hsp40 with affinity for phosphatidic acid lipids

Zhang

Young

Foster

et al. 2021

Preprint

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“…Overexpressing MET13 significantly improved the expression of RuCelA. Caj1 is a chaperone that regulates the stability or turnover of plasma membrane proteins [ 45 ]. Out of the 102 ESRTs overexpressed in this study, CAJ1 was the only gene that improved the secretory expressions of both AnFaeA and RuCelA.…”

Section: Resultsmentioning

confidence: 99%

Characterization and modulation of endoplasmic reticulum stress response target genes in Kluyveromyces marxianus to improve secretory expressions of heterologous proteins

Shi

Zhou

Xue

et al. 2021

Biotechnol Biofuels

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Background Kluyveromyces marxianus is a promising cell factory for producing bioethanol and that raised a demand for a high yield of heterologous proteins in this species. Expressions of heterologous proteins usually lead to the accumulation of misfolded or unfolded proteins in the lumen of the endoplasmic reticulum (ER) and then cause ER stress. To cope with this problem, a group of ER stress response target genes (ESRTs) are induced, mainly through a signaling network called unfolded protein response (UPR). Characterization and modulation of ESRTs direct the optimization of heterologous expressions. However, ESRTs in K. marxianus have not been identified so far. Results In this study, we characterized the ER stress response in K. marxianus for the first time, by using two ER stress-inducing reagents, dithiothreitol (DTT) and tunicamycin (TM). Results showed that the Kar2–Ire1–Hac1 pathway of UPR is well conserved in K. marxianus. About 15% and 6% of genes were upregulated during treatment of DTT and TM, respectively. A total of 115 upregulated genes were characterized as ESRTs, among which 97 genes were identified as UPR target genes and 37 UPR target genes contained UPR elements in their promoters. Genes related to carbohydrate metabolic process and actin filament organization were identified as new types of UPR target genes. A total of 102 ESRTs were overexpressed separately in plasmids and their effects on productions of two different lignocellulolytic enzymes were systematically evaluated. Overexpressing genes involved in carbohydrate metabolism, including PDC1, PGK and VID28, overexpressing a chaperone gene CAJ1 or overexpressing a reductase gene MET13 substantially improved secretion expressions of heterologous proteins. Meanwhile, overexpressing a novel gene, KLMA_50479 (named ESR1), as well as overexpressing genes involved in ER-associated protein degradation (ERAD), including HRD3, USA1 andYET3, reduced the secretory expressions. ESR1 and the aforementioned ERAD genes were deleted from the genome. Resultant mutants, except the yet3Δ mutant, substantially improved secretions of three different heterologous proteins. During the fed-batch fermentation, extracellular activities of an endoxylanase and a glucanase in hrd3Δ cells improved by 43% and 28%, respectively, compared to those in wild-type cells. Conclusions Our results unveil the transcriptional scope of the ER stress response in K. marxianus and suggest efficient ways to improve productions of heterologous proteins by manipulating expressions of ESRTs.

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Dosage sensitivity of JDPs, a valuable tool for understanding their function: a case study on Caj1 overexpression-mediated filamentous growth in budding yeast

2021

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Over-expression of Caj1, a plasma membrane associated J-domain protein in Saccharomyces cerevisiae, stabilizes amino acid permeases

Cited by 6 publications

References 57 publications

Nanodiscs-based proteomics identify Caj1 as an Hsp40 with affinity for phosphatidic acid lipids

Nanodiscs-based proteomics identify Caj1 as an Hsp40 with affinity for phosphatidic acid lipids

Characterization and modulation of endoplasmic reticulum stress response target genes in Kluyveromyces marxianus to improve secretory expressions of heterologous proteins

Dosage sensitivity of JDPs, a valuable tool for understanding their function: a case study on Caj1 overexpression-mediated filamentous growth in budding yeast

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