Distinct AAA-ATPase p97 complexes function in discrete steps of nuclear assembly

Abstract

Although nuclear envelope (NE) assembly is known to require the GTPase Ran, the membrane fusion machinery involved is uncharacterized. NE assembly involves formation of a reticular network on chromatin, fusion of this network into a closed NE and subsequent expansion. Here we show that p97, an AAA-ATPase previously implicated in fusion of Golgi and transitional endoplasmic reticulum (ER) membranes together with the adaptor p47, has two discrete functions in NE assembly. Formation of a closed NE requires the p97–Ufd1–Npl4 complex, not previously implicated in membrane fusion. Subsequent NE growth involves a p97–p47 complex. This study provides the first insights into the molecular mechanisms and specificity of fusion events involved in NE formation.

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Figure 1: Formation of NE and ER in vitro.
Figure 2: p97 is involved in NE and ER formation.
Figure 3: Depletion of p97 adaptors.
Figure 4: Different functions for Ufd1–Npl4 and p47.
Figure 5: Schematic step-wise model of NE formation.

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Acknowledgements

M.H. was supported by EMBO and H.H.M. by the Human Frontiers Science Programme Organisation. We thank Jan Ellenberg and the ELMF for help with microscopy. Thanks to W. Antonin, P. Askjaer, K. Czaplinski, L. Englmeier, J. Ellenberg, V. Hachet, B. Huelsmann, M. Ohno and C. Schatz for critical comments on the manuscript.

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Correspondence to Iain W. Mattaj.

Supplementary information

Supplementary figures

Figure S1 Nuclei formed in the presence of p97 are functional. (PDF 348 kb)

Figure S2 Characterization of NE proteins in p97-restored nuclei.

Figure S3 Nuclei formed in the absence of p47 have an intact NE.

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Hetzer, M., Meyer, H., Walther, T. et al. Distinct AAA-ATPase p97 complexes function in discrete steps of nuclear assembly. Nat Cell Biol 3, 1086–1091 (2001). https://doi.org/10.1038/ncb1201-1086

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