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

Sagarika, Preeti; Dobriyal, Neha; Sahi, Chandan

doi:10.1007/s00294-021-01153-8

Cited by 4 publications

(7 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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. , 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 . 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: 68%

“…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. ,, Besides Caj1, we also identify the phosphatidylinositol (PI)-interacting proteins Pdr16, Pik1, and Inp53. Pdr16 is annotated as a phosphatidylinositol transfer protein; Pik1 is a phosphatidylinositol kinase, and Inp53 is a phosphatidylinositol phosphatase.…”

Section: Resultsmentioning

confidence: 96%

See 1 more Smart Citation

Nanodisc-Based Proteomics Identify Caj1 as an Hsp40 with Affinity for Phosphatidic Acid Lipids

et al. 2021

View full text Add to dashboard Cite

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 a lipid bilayer encircled with amphipathic scaffold proteinsalong with quantitative proteomics to identify lipid-binding proteins in Saccharomyces 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 (PEPAnanodiscs), 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 the 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 combining proteomics with lipid nanodiscs to identify and characterize protein−lipid interactions that may not be evident using other methods. Data are available via ProteomeXchange with the identifier PXD027992.

show abstract

Section: Introductionsupporting

confidence: 68%

Section: Resultsmentioning

confidence: 96%

Nanodisc-Based Proteomics Identify Caj1 as an Hsp40 with Affinity for Phosphatidic Acid Lipids

et al. 2021

View full text Add to dashboard Cite

show abstract

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%

“…What is the function of Caj1 in vivo ? 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 .…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

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

Zhang

Young

Foster

et al. 2021

Preprint

View full text Add to dashboard Cite

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.

show abstract

J-like protein family of Arabidopsis thaliana: the enigmatic cousins of J-domain proteins

et al. 2022

View full text Add to dashboard Cite

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

Cited by 4 publications

References 61 publications

Nanodisc-Based Proteomics Identify Caj1 as an Hsp40 with Affinity for Phosphatidic Acid Lipids

Nanodisc-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

J-like protein family of Arabidopsis thaliana: the enigmatic cousins of J-domain proteins

Contact Info

Product

Resources

About