Fatty acid synthase cooperates with protrudin to facilitate membrane outgrowth of cellular protrusions
نویسندگان
چکیده
Cellular protrusion formation capacity is a key feature of developing neurons and many eukaryotic cells. However, the mechanisms underlying membrane growth in protrusion formation are largely unclear. In this study, photo-reactive unnatural amino acid 3-(3-methyl-3H-diazirin-3-yl)-propamino-carbonyl-Nε-l-lysine was incorporated by a genetic code expansion strategy into protrudin, a protein localized in acidic endosomes and in the endoplasmic reticulum, that induces cellular protrusion and neurite formation. The modified protrudin was used for covalent trapping of protrudin-interacting proteins in living cells. Fatty acid synthase (FASN), which synthesizes free fatty acids, was identified to transiently interact with protrudin. Further characterization revealed a unique cooperation mechanism in which protrudin cooperates with FASN to facilitate cellular protrusion formation. This work reveals a novel mechanism involved in protrusion formation that is dependent on transient interaction between FASN and protrudin, and establishes a creative strategy to investigate transient protein-protein interactions in mammalian cells.
منابع مشابه
Phosphoinositides differentially regulate protrudin localization through the FYVE domain.
Protrudin is a FYVE (Fab 1, YOTB, Vac 1, and EEA1) domain-containing protein involved in transport of neuronal cargoes and implicated in the onset of hereditary spastic paraplegia. Our image-based screening of the lipid binding domain library revealed novel plasma membrane localization of the FYVE domain of protrudin unlike canonical FYVE domains that are localized to early endosomes. The membr...
متن کاملA grab to move on: ER-endosome contacts in membrane protrusion formation and neurite outgrowth.
A key feature of many eukaryotic cells, most prominently seen in developing neurons, is their ability to form and extend membrane protrusions. How protrusion formation is linked to exocytic membrane trafficking is largely unclear. In a recent paper published in Nature, Raiborg et al identify a crucial role in this process for dynamic membrane contact sites (MCSs) between the ER and endosomes. T...
متن کاملHave you seen? A grab to move on: ER–endosome contacts in membrane protrusion formation and neurite outgrowth
A key feature of many eukaryotic cells, most prominently seen in developing neurons, is their ability to form and extend membrane protrusions. How protrusion formation is linked to exocytic membrane trafficking is largely unclear. In a recent paper published in Nature, Raiborg et al identify a crucial role in this process for dynamic membrane contact sites (MCSs) between the ER and endosomes. T...
متن کاملProtrudin serves as an adaptor molecule that connects KIF5 and its cargoes in vesicular transport during process formation
Neurons are highly polarized cells with long neurites. Vesicular transport is required for neurite extension. We recently identified protrudin as a key regulator of vesicular transport during neurite extension. Expression of protrudin in nonneuronal cells thus induces formation of neurite-like membrane protrusions. We adopted a proteomics approach to identify proteins that associate with protru...
متن کاملPromotion of neurite extension by protrudin requires its interaction with vesicle-associated membrane protein-associated protein.
Protrudin is a protein that contains a Rab11-binding domain and a FYVE (lipid-binding) domain and that functions to promote neurite formation through interaction with the GDP-bound form of Rab11. Protrudin also contains a short sequence motif designated FFAT (two phenylalanines in an acidic tract), which in other proteins has been shown to mediate binding to vesicle-associated membrane protein-...
متن کامل