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Spatially regulated ubiquitin ligation by an ER/nuclear membrane ligase


The ubiquitin system targets many cellular proteins. Doa10 (also known as Ssm4), a yeast transmembrane ubiquitin ligase (E3), resides in the endoplasmic reticulum (ER), but it attaches ubiquitin to soluble proteins that concentrate in the nucleus. A central question is how nuclear substrates gain access to an enzyme in the ER. Here we show that Doa10 reaches the inner nuclear membrane. A subcomplex of nuclear pore subunits is important for this transport. Notably, another ER transmembrane E3, Hrd1 (also known as Der3), cannot localize efficiently to the inner nuclear membrane. Tethering Doa10 at the cell periphery inhibits degradation of soluble nuclear substrates but not cytoplasmic ones. If Doa10 is released from these peripheral sites, localization of Doa10 to the nuclear envelope and degradation of its nuclear substrates are restored in parallel. Thus, localization of Doa10 to the inner nuclear membrane is necessary for nuclear substrate degradation. These data indicate that different membrane ubiquitin ligases are spatially sorted within the ER–nuclear envelope membrane system and that this differential localization contributes to their specificity.

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Figure 1: Localization of a transmembrane ubiquitin ligase to the inner nuclear envelope.
Figure 2: Fluorescence assay of INM protein localization.
Figure 3: Localization of Doa10 to the INM is required for nuclear substrate degradation.
Figure 4: Nuclear substrate degradation and determinants of Doa10 INM localization.


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We thank R. Sternglanz and R. Wozniak for strains and plasmids, P. Crews for generously providing LTA, O. Kerscher for Supplementary Fig. 1, and J. Wu and J. Wolenski for advice on confocal microscopy. We are grateful to S. Kreft, A. Lewis and T. Ravid for comments on the manuscript. This work was supported by the NIH. Author Contributions M.D. performed all the experiments, and M.D. and M.H. planned the experiments, performed the data analysis and wrote the paper.

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Correspondence to Mark Hochstrasser.

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Deng, M., Hochstrasser, M. Spatially regulated ubiquitin ligation by an ER/nuclear membrane ligase. Nature 443, 827–831 (2006).

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