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SNX27–Retromer directly binds ESCPE-1 to transfer cargo proteins during endosomal recycling
Abstract
Coat complexes coordinate cargo recognition through cargo adaptors with biogenesis of transport carriers during integral membrane protein trafficking. Here, we combine biochemical, structural, and cellular analyses to establish the mechanistic basis through which SNX27–Retromer, a major endosomal cargo adaptor, couples to the membrane remodeling endosomal SNX-BAR sorting complex for promoting exit 1 (ESCPE-1). In showing that the SNX27 FERM (4.1/ezrin/radixin/moesin) domain directly binds acidic-Asp-Leu-Phe (aDLF) motifs in the SNX1/SNX2 subunits of ESCPE-1, we propose a handover model where SNX27–Retromer captured cargo proteins are transferred into ESCPE-1 transport carriers to promote endosome-to-plasma membrane recycling. By revealing that assembly of the SNX27:Retromer:ESCPE-1 coat evolved in a stepwise manner during early metazoan evolution, likely reflecting the increasing complexity of endosome-to-plasma membrane recycling from the ancestral opisthokont to modern animals, we provide further evidence of the functional diversification of yeast pentameric Retromer in the recycling of hundreds of integral membrane proteins in metazoans.
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VPS35 suppression resulted in an increase in LRFNs (LRFN1, 2 and 4) at the neuronal surface, suggesting that SNX27 can act independently to Retromer in the trafficking of a sub-set of cargoes, potentially through the ESCPE-1 complex.
“Surprisingly, VPS35 suppression resulted in an increase in LRFNs (LRFN1, 2 and 4) at the neuronal surface (Figure 2C) suggesting that SNX27 can act independently to Retromer in the trafficking of a sub-set of cargoes, potentially through the ESCPE-1 complex [27,28].”
From Global identification of neuronal and astrocytic integral membrane proteins that require Retromer for their endosomal recycling (2026) · cites this paper as [27]
The Results section reports that SNX27 binds ESCPE-1 even when SNX27 mutants cannot bind Retromer, supporting a Retromer-independent physical interaction. It does not establish Retromer-independent trafficking of a cargo subset; the Discussion instead describes Retromer as essential for retrieval of the cargos studied. The neuronal LRFN surface increase is presented as the citing authors’ own Figure 2C result, not as a finding from this source. (LLM, conf=0.84)
Experimental — accuracy not yet measuredRetromer associates with accessory proteins and cargo adaptors including sorting nexins, the WASH complex, RAB GTPase regulators including TBC1D5 and ANKRD27/VARP, and RME8.
“To function, Retromer associates with accessory proteins and cargo adaptors including sorting nexins (SNXs) [24][25][26][27][28][29][30][31][32][33][34], the WASH complex [35][36][37][38][39], RAB GTPase regulators including TBC1D5 and ANKRD27/VARP [40][41][42][43][44][45][46], and RME8 [47].”
From Global identification of neuronal and astrocytic integral membrane proteins that require Retromer for their endosomal recycling (2026) · cites this paper as [27]
SNX27 interacts with SNX-BAR proteins SNX1 and SNX2 and the WASH complex to couple sequence-dependent retrieval of SNX27 cargoes with tubule formation during cargo recycling.
“SNX27 also interacts with the SNX-BAR (Bin/Amphiphysin/Rvs) proteins SNX1 and SNX2 [27,28,31,62] and the WASH complex [37] to couple sequence-dependent retrieval of SNX27 cargoes with tubule formation during the process of cargo recycling.”
From Global identification of neuronal and astrocytic integral membrane proteins that require Retromer for their endosomal recycling (2026) · cites this paper as [27]
The lack of significantly enriched labelling between SNX27 and Retromer may result from a technical limitation or reflect a highly dynamic, transient association between cargo-loaded SNX27 and the Retromer demarcated sub-domain during the handover of captured cargo.
“Given the direct nature of the PDZ domain of SNX27 in binding to VPS26A and VPS26B and the importance of this coupling for Retromer-mediated cargo sorting11, the lack of significantly enriched labelling may result from a technical limitation or reflect a highly dynamic, transient association between cargo-loaded SNX27 and the Retromer demarcated sub-domain during the handover of captured cargo31.”
From Mapping of endosomal proximity proteomes reveals Retromer as a hub for RAB GTPase regulation (2025) · cites this paper as [31]
ID1-SNX27 also labelled ESCPE-1 subunits, supporting the recently proposed cargo handover model for SNX27-Retromer mediated ESCPE-1-dependent cargo sorting.
“In addition, and consistent with the colocalization of endogenous VPS35, SNX27 and SNX1 (Fig. 2I and Supplementary Fig. 4D), ID1-SNX27 also labelled ESCPE-1 subunits (Fig. 2E), supporting the recently proposed cargo handover model for SNX27-Retromer mediated ESCPE-1-dependent cargo sorting31.”
From Mapping of endosomal proximity proteomes reveals Retromer as a hub for RAB GTPase regulation (2025) · cites this paper as [31]
VPS35KO, SNX27KO, SNX17KO, and VPS35LKO clonal cell lines used in this study were previously characterised.
“VPS35KO, SNX27KO, SNX17KO, and VPS35LKO clonal cell lines used in this study were previously characterised19,20,31.”
From Mapping of endosomal proximity proteomes reveals Retromer as a hub for RAB GTPase regulation (2025) · cites this paper as [31]
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Citations
Cites
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ColabFold - Making protein folding accessible to all
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“(A) Alphafold2 structural prediction for FL human SNX27 [44].”
BackgroundThe Genome of the Ctenophore Mnemiopsis leidyi and Its Implications for Cell Type Evolution
Ryan, J. F., Pang, K., Schnitzler, C. E., Nguyen, A.-D., Moreland, R. T., Simmons, D. K., Koch, B. J., Francis, W. R., Havlak, P., Program, N. C. S., et al. (2013) · Science · doi:10.1126/science.1242592 · Cited by 682
Retriever is a multiprotein complex for retromer-independent endosomal cargo recycling
McNally, K. E., Faulkner, R., Steinberg, F., Gallon, M., Ghai, R., Pim, D., Langton, P., Pearson, N., Danson, C. M., Nägele, H., et al. (2017) · Nature Cell Biology · doi:10.1038/ncb3610 · Cited by 346
Self-citationCCC- and WASH-mediated endosomal sorting of LDLR is required for normal clearance of circulating LDL
Bartuzi, P., Billadeau, D. D., Favier, R., Rong, S., Dekker, D., Fedoseienko, A., Fieten, H., Wijers, M., Levels, J. H., Huijkman, N., et al. (2016) · Nature Communications · doi:10.1038/ncomms10961 · Cited by 205
Endosomal retrieval of cargo: Retromer is not alone
McNally, K. E., & Cullen, P. J. (2018) · Trends in Cell Biology · doi:10.1016/j.tcb.2018.06.005 · Cited by 169
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Metadata
- Authors
- Simonetti, B., Guo, Q., Giménez-Andrés, M., Chen, K.-E., Moody, E. R. R., Evans, A. J., Chandra, M., Danson, C. M., Williams, T. A., Collins, B. M., & Cullen, P. J.
- Venue
- PLOS Biology
- Publisher
- Public Library of Science (PLoS)
- Published
- 2022-04-13
- Volume
- 20
- Issue
- 4
- Pages
- e3001601
- ISSN
- 1545-7885
- PMC
- PMC9038204
- Cited by
- 55
- References
- 7
- Language
- en
- Update notices
- new_version (2022-04-25)
- Funders
- Wellcome Trust (104568/Z/14/Z, 220260/Z/20/Z)
- Medical Research Council (MR/L007363/1, MR/P018807/1)
- Lister Institute of Preventive Medicine
- Royal Society Noreen Murray Research Professorship (RSRP/R1/211004)
- National Health and Medical Research Council (APP1136021, APP1156493)
- Research Fellows Enhancement Award (RGF\EA\180199)
- Royal Society University Fellowship (URF\R\201024)
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