1. Disrupted ER membrane protein complex-mediated topogenesis drives congenital neural crest defects
- Author
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Marquez, Jonathan, Criscione, June, Charney, Rebekah M., Prasad, Maneeshi S., Hwang, Woong Y., Mis, Emily K., Garcia-Castro, Martin I., and Khokha, Mustafa K.
- Subjects
Thermo Fisher Scientific Inc. ,Yale University. School of Medicine ,Congenital heart defects ,Heart diseases ,Medical research ,Cardiac patients ,Genetic disorders ,Membrane proteins ,Scientific equipment industry ,Proteomics ,Phenotypes ,Diseases ,Birth defects ,Proteins ,Cardiovascular diseases ,Alleles - Abstract
Introduction Multipass membrane proteins cross lipid bilayers multiple times with specific topologies essential for their function. The multipass membrane proteins that contain a lumenal N-terminus require the endoplasmic reticulum (ER) [...], Multipass membrane proteins have a myriad of functions, including transduction of cell-cell signals, ion transport, and photoreception. Insertion of these proteins into the membrane depends on the endoplasmic reticulum (ER) membrane protein complex (EMC). Recently, birth defects have been observed in patients with variants in the gene encoding a member of this complex, EMC1. Patient phenotypes include congenital heart disease, craniofacial malformations, and neurodevelopmental disease. However, a molecular connection between EMC1 and these birth defects is lacking. Using Xenopus, we identified defects in neural crest cells (NCCs) upon emcl depletion. We then used unbiased proteomics and discovered a critical role for emcl in WNT signaling. Consistent with this, readouts of WNT signaling and Frizzled (Fzd) levels were reduced in emcl-depleted embryos, while NCC defects could be rescued with [beta]-catenin. Interestingly, other transmembrane proteins were mislocalized upon emcl depletion, providing insight into additional patient phenotypes. To translate our findings back to humans, we found that EMC1 was necessary for human NCC development in vitro. Finally, we tested patient variants in our Xenopus model and found the majority to be loss-of-function alleles. Our findings define molecular mechanisms whereby EMC1 dysfunction causes disease phenotypes through dysfunctional multipass membrane protein topogenesis.
- Published
- 2020
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