PUBLICATION
The human nephrin Y1139RSL motif is essential for podocyte foot process organization and slit diaphragm formation during glomerular development
- Authors
- Espiritu, E.B., Jiang, H., Moreau-Marquis, S., Sullivan, M., Yan, K., Beer Stolz, D., Sampson, M.G., Hukriede, N.A., Swiatecka-Urban, A.
- ID
- ZDB-PUB-190604-6
- Date
- 2019
- Source
- The Journal of biological chemistry 294(28): 10773-10788 (Journal)
- Registered Authors
- Hukriede, Neil
- Keywords
- cathrin-dependent endocytosis, cell adhesion, cell culture, cell surface protein, cell-cell interaction, clathrin, endocytosis, foot processes, nephrin, nephrology, nephrotic syndrome, podocyte, slit diaphragm, zebrafish
- MeSH Terms
-
- Amino Acid Motifs
- Animals
- Cell Line
- Cell Membrane/metabolism
- Cell Movement
- Clathrin/metabolism
- Embryo, Nonmammalian/metabolism
- Endocytosis
- Humans
- Kidney Glomerulus/metabolism*
- Kidney Glomerulus/ultrastructure
- Membrane Proteins/antagonists & inhibitors
- Membrane Proteins/genetics
- Membrane Proteins/metabolism*
- Morpholinos/metabolism
- Mutagenesis, Site-Directed
- Phosphorylation
- Podocytes/cytology
- Podocytes/metabolism
- Zebrafish/growth & development
- PubMed
- 31152064 Full text @ J. Biol. Chem.
Citation
Espiritu, E.B., Jiang, H., Moreau-Marquis, S., Sullivan, M., Yan, K., Beer Stolz, D., Sampson, M.G., Hukriede, N.A., Swiatecka-Urban, A. (2019) The human nephrin Y1139RSL motif is essential for podocyte foot process organization and slit diaphragm formation during glomerular development. The Journal of biological chemistry. 294(28):10773-10788.
Abstract
Nephrin is an immunoglobulin-type cell-adhesion molecule with a key role in the glomerular inter-podocyte slit diaphragm. Mutations in the nephrin gene are associated with defects in the slit diaphragm, leading to early-onset nephrotic syndrome, typically resistant to treatment. Although the endocytic trafficking of nephrin is essential for the assembly of the slit diaphragm, nephrin's specific endocytic motifs remain unknown. To search for endocytic motifs, here we performed a multisequence alignment of nephrin and identified a canonical YxxΦ-type motif, Y1139RSL, in the nephrin cytoplasmic tail, expressed only in primates. Using site-directed mutagenesis, various biochemical methods, single-plane illumination microscopy (SPIM), a human podocyte line, and a human nephrin-expressing zebrafish model, we found that Y1139RSL is a novel endocytic motif and a structural element for clathrin-mediated nephrin endocytosis that functions as a phosphorylation-sensitive signal. We observed that Y1139RSL motif-mediated endocytosis helps to localize nephrin to specialized plasma membrane domains in podocytes and is essential for normal foot process organization into a functional slit diaphragm between neighboring foot processes in zebrafish. The importance of nephrin Y1139RSL for healthy podocyte development was supported by population-level analyses of genetic variations at this motif, revealing that such variations are very rare, suggesting that mutations in this motif haveautosomal-recessive negative effects on kidney health. These findings expand our understanding of the mechanism underlying nephrin endocytosis and may lead to improved diagnostic tools or therapeutic strategies for managing early-onset, treatment-resistant nephrotic syndrome.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping