ATPase activity of DFCP1 controls selective autophagy

Nähse, V.; Raiborg, C.; Tan, K. W.; Mørk, S.; Torgersen, M. L.; Wenzel, E. M.; Nager, M.; Salo, V. T.; Johansen, T.; Ikonen, E.; Schink, K. O.; Stenmark, H.

doi:10.1101/2022.02.24.481614

bioRxiv · 10.1101/2022.02.24.481614

ATPase activity of DFCP1 controls selective autophagy

Nähse, V.·Raiborg, C.·Tan, K. W.·Mørk, S.·Torgersen, M. L.·Wenzel, E. M.·Nager, M.·Salo, V. T.·Johansen, T.·Ikonen, E.·Schink, K. O.·Stenmark, H.

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Abstract

Cellular homeostasis is governed by removal of damaged organelles and protein aggregates by selective autophagy mediated by cargo adaptors such as p62/SQSTM1. Autophagosomes can assemble in specialized cup-shaped regions of the endoplasmic reticulum (ER) known as omegasomes, which are characterized by the presence of the ER protein DFCP1/ZFYVE1. The function of DFCP1 is unknown, as are the mechanisms of omegasome formation and constriction. Here, we demonstrate that DFCP1 is an ATPase that dimerizes in an ATP-dependent fashion. Whereas depletion of DFCP1 had a minor effect on bulk autophagic flux, DFCP1 was required to maintain the autophagic flux of p62 under both fed and starved conditions, and this was dependent on its ability to bind and hydrolyse ATP. While DFCP1 mutants defective in ATP binding or hydrolysis localized to forming omegasomes, these omegasomes failed to constrict. Consequently, the release of nascent autophagosomes from omegasomes was markedly delayed. DFCP1 was found to associate with ubiquitinated proteins, and degradation of ubiquitinated cargoes such as protein aggregates, mitochondria and micronuclei was strongly inhibited when DFCP1 was knocked out or mutated. Thus, DFCP1 mediates ATPase-driven constriction of omegasomes to release autophagosomes for selective autophagy.

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Nähse, V., Raiborg, C., Tan, K. W., Mørk, S., Torgersen, M. L., Wenzel, E. M., Nager, M., Salo, V. T., Johansen, T., Ikonen, E., Schink, K. O., Stenmark, H.. 2022-02-24. ATPase activity of DFCP1 controls selective autophagy. https://doi.org/10.1101/2022.02.24.481614

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