ATPase activity of DFCP1 controls selective autophagy

N&aumlhse, Viola; Raiborg, Camilla; Tan, Kia Wee; M&oslashrk, Sissel; Torgersen, Maria Lyngaas; Wenzel, Eva M.; Nàger, Mireia; Salo, Veijo T.; Johansen, Terje; Ikonen, Elina; Schink, Kay Oliver; Stenmark, Harald

doi:10.1101/2022.02.24.481614

Cited by 8 publications

(21 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Upon starvation, DFCP1 accumulates in punctate structures in proximity of the ER. These DFCP1-positive structures grow and form rings around cargo that collapse back into a punctate structure before completely disappearing ( 9 , 38 ). To capture early events during phagophore formation, we overexpressed mCherry-tagged DFCP1 and recorded 25 tomograms at sites of colocalization with the GFP- Salmonella signal.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

In situ snapshots along a mammalian selective autophagy pathway

Tripathi-Giesgen

Schulman

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Selective macroautophagy (hereafter referred to as autophagy) describes a process in which cytosolic material is engulfed in a double membrane organelle called an autophagosome. Autophagosomes are carriers responsible for delivering their content to a lytic compartment for destruction. The cargo can be of diverse origin, ranging from macromolecular complexes to protein aggregates, organelles, and even invading pathogens. Each cargo is unique in composition and size, presenting different challenges to autophagosome biogenesis. Among the largest cargoes targeted by the autophagy machinery are intracellular bacteria, which can, in the case of Salmonella, range from 2 to 5 μm in length and 0.5 to 1.5 μm in width. How phagophores form and expand on such a large cargo remains mechanistically unclear. Here, we used HeLa cells infected with an auxotrophic Salmonella to study the process of phagophore biogenesis using in situ correlative cryo-ET. We show that host cells generate multiple phagophores at the site of damaged Salmonella -containing vacuoles (SCVs). The observed double membrane structures range from disk-shaped to expanded cup-shaped phagophores, which have a thin intermembrane lumen with a dilating rim region and expand using the SCV, the outer membrane of Salmonella , or existing phagophores as templates. Phagophore rims establish different forms of contact with the endoplasmic reticulum (ER) via structurally distinct molecular entities for membrane formation and expansion. Early omegasomes correlated with the marker Double-FYVE domain-Containing Protein 1 (DFCP1) are observed in close association with the ER without apparent membrane continuity. Our study provides insights into the formation of phagophores around one of the largest selective cargoes.

show abstract

Section: Resultsmentioning

confidence: 99%

“…Recently, DFCP1 has been shown to be an ATPase important for omegasome constriction ( 38 ). The similarities of DFCP1 to the well-studied GTPase dynamin suggest that the observed filaments at the contact sites with the ER may represent DFCP1 assemblies.…”

Section: Discussionmentioning

confidence: 99%

In situ snapshots along a mammalian selective autophagy pathway

Tripathi-Giesgen

Schulman

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…WIPI2 is necessary for recruiting the lipidation machinery for the phagophore-forming protein LC3B (45,46), whereas the ATPase DFCP1 is necessary for releasing the newly formed autophagosomes from the ER (47). Despite the reduction of WIPI2 and DFCP1 recruitment upon autophagy induction in LRBA-KO cells, comparable levels of LC3 lipidation to WT cells were observed.…”

Section: Discussionmentioning

confidence: 92%

LRBA balances antigen presentation and T-cell responsesviaautophagy by binding to PIK3R4 and FYCO1

Gámez-Díaz

Ligeon

Sindram

et al. 2022

Preprint

View full text Add to dashboard Cite

Patients with lipopolysaccharide responsive beige-like anchor protein (LRBA) deficiency have an exacerbated T-cell activation and a defective B-cell response. Previously, we suggested reduced autophagy as the biological cause of the aberrant humoral response; however, the exact molecular mechanism of LRBA in autophagy and its impact on T-cell responses remain unknown. We screened for LRBA-interacting proteins, finding that LRBA interacts with the phosphoinositide 3-kinase regulatory subunit 4 (PIK3R4) and with the FYVE And Coiled-Coil Domain Autophagy Adaptor 1 (FYCO1). Interestingly, both proteins play essential roles at different stages of autophagy. Specifically, PIK3R4 facilitates the production of phosphatidylinositol-3 phosphate (PI(3)P) required for the recruitment of PI(3)P-binding adaptor proteins allowing autophagosome formation and autophagosome-lysosome fusion, whereas FYCO1 allows autophagosome movement. LRBA-KO cells showed an impaired production of PI(3)P, a delayed autophagosome-lysosome fusion, an accumulation of abnormal autophagosomes and an atypical lysosomal positioning. These observations explain the decreased cargo material degradation and the overall defective autophagy flux in LRBA-KO cells. Abnormal autophagosomes in LRBA-KO cells are associated with prolonged antigen presentation to T cells, resulting in a higher production of proinflammatory cytokines, as autophagy is a major intracellular degradation system for major histocompatibility class II complex (MHCII) loading. Taken together, our data suggest that i) LRBA forms different protein complexes serving at different stages of autophagy, and ii) loss of LRBA impacts the targeting of cytosolic antigens for autophagy degradation, enhancing antigen presentation. The latter could contribute to the exacerbated T-cell immune dysregulation observed in LRBA-deficient patients

show abstract

“…[ 77 ] A recent study has shown that DFCP1 (Double FYVE‐Containing Protein 1), a protein involved in forming initial membrane structures for autophagosomes, [ 78,79 ] possesses ATPase activity. [ 80 ] The need for energy in autophagy is further supported by recent research demonstrating that functional mitochondria are necessary for autophagy induction [ 22,81 ] or for maximizing autophagy. [ 82 ] This emphasizes the crucial role of mitochondria in facilitating autophagy initiation.…”

Section: Autophagy Needs Energy or Functional Mitochondriamentioning

confidence: 99%

Contrasting views on the role of AMPK in autophagy

Kim

2024

BioEssays

View full text Add to dashboard Cite

Efficient management of low energy states is vital for cells to maintain basic functions and metabolism and avoid cell death. While autophagy has long been considered a critical mechanism for ensuring survival during energy depletion, recent research has presented conflicting evidence, challenging the long‐standing concept. This recent development suggests that cells prioritize preserving essential cellular components while restraining autophagy induction when cellular energy is limited. This essay explores the conceptual discourse on autophagy regulation during energy stress, navigating through the studies that established the current paradigm and the recent research that has challenged its validity while proposing an alternative model. This exploration highlights the far‐reaching implications of the alternative model, which represents a conceptual departure from the established paradigm, offering new perspectives on how cells respond to energy stress.

show abstract

ATPase activity of DFCP1 controls selective autophagy

Cited by 8 publications

References 46 publications

In situ snapshots along a mammalian selective autophagy pathway

In situ snapshots along a mammalian selective autophagy pathway

LRBA balances antigen presentation and T-cell responsesviaautophagy by binding to PIK3R4 and FYCO1

Contrasting views on the role of AMPK in autophagy

Contact Info

Product

Resources

About