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Linkage reprogramming by tailor-made E3s reveals polyubiquitin chain requirements in DNA-damage bypass

Sabrina Wegmann,Cindy Meister,Christian Renz,George Yakoub,Hans-Peter Wollscheid,Diane T. Takahashi,Ivan Mikicic,Petra Beli,H. Ulrich

Published 2022 in Molecules and Cells

ABSTRACT

Summary A polyubiquitin chain can adopt a variety of shapes, depending on how the ubiquitin monomers are joined. However, the relevance of linkage for the signaling functions of polyubiquitin chains is often poorly understood because of our inability to control or manipulate this parameter in vivo. Here, we present a strategy for reprogramming polyubiquitin chain linkage by means of tailor-made, linkage- and substrate-selective ubiquitin ligases. Using the polyubiquitylation of the budding yeast replication factor PCNA in response to DNA damage as a model case, we show that altering the features of a polyubiquitin chain in vivo can change the fate of the modified substrate. We also provide evidence for redundancy between distinct but structurally similar linkages, and we demonstrate by proof-of-principle experiments that the method can be generalized to targets beyond PCNA. Our study illustrates a promising approach toward the in vivo analysis of polyubiquitin signaling.

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