K27-linked ubiquitylation promotes p97 substrate processing and is essential for cell proliferation
Research output: Contribution to journal › Journal article › Research › peer-review
Standard
K27-linked ubiquitylation promotes p97 substrate processing and is essential for cell proliferation. / Shearer, Robert F; Typas, Dimitris; Coscia, Fabian; Schovsbo, Sofie; Kruse, Thomas; Mund, Andreas; Mailand, Niels.
In: E M B O Journal, Vol. 41, No. 9, e110145, 2022.Research output: Contribution to journal › Journal article › Research › peer-review
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - K27-linked ubiquitylation promotes p97 substrate processing and is essential for cell proliferation
AU - Shearer, Robert F
AU - Typas, Dimitris
AU - Coscia, Fabian
AU - Schovsbo, Sofie
AU - Kruse, Thomas
AU - Mund, Andreas
AU - Mailand, Niels
N1 - © 2022 The Authors. Published under the terms of the CC BY NC ND 4.0 license.
PY - 2022
Y1 - 2022
N2 - Conjugation of ubiquitin (Ub) to numerous substrate proteins regulates virtually all cellular processes. Eight distinct ubiquitin polymer linkages specifying different functional outcomes are generated in cells. However, the roles of some atypical poly-ubiquitin topologies, in particular linkages via lysine 27 (K27), remain poorly understood due to a lack of tools for their specific detection and manipulation. Here, we adapted a cell-based ubiquitin replacement strategy to enable selective and conditional abrogation of K27-linked ubiquitylation, revealing that this ubiquitin linkage type is essential for proliferation of human cells. We demonstrate that K27-linked ubiquitylation is predominantly a nuclear modification whose ablation deregulates nuclear ubiquitylation dynamics and impairs cell cycle progression in an epistatic manner with inactivation of the ATPase p97/VCP. Moreover, we show that a p97-proteasome pathway model substrate (Ub(G76V)-GFP) is directly modified by K27-linked ubiquitylation, and that disabling the formation of K27-linked ubiquitin signals or blocking their decoding via overexpression of the K27 linkage-specific binder UCHL3 impedes Ub(G76V)-GFP turnover at the level of p97 function. Our findings suggest a critical role of K27-linked ubiquitylation in supporting cell fitness by facilitating p97-dependent processing of ubiquitylated nuclear proteins.
AB - Conjugation of ubiquitin (Ub) to numerous substrate proteins regulates virtually all cellular processes. Eight distinct ubiquitin polymer linkages specifying different functional outcomes are generated in cells. However, the roles of some atypical poly-ubiquitin topologies, in particular linkages via lysine 27 (K27), remain poorly understood due to a lack of tools for their specific detection and manipulation. Here, we adapted a cell-based ubiquitin replacement strategy to enable selective and conditional abrogation of K27-linked ubiquitylation, revealing that this ubiquitin linkage type is essential for proliferation of human cells. We demonstrate that K27-linked ubiquitylation is predominantly a nuclear modification whose ablation deregulates nuclear ubiquitylation dynamics and impairs cell cycle progression in an epistatic manner with inactivation of the ATPase p97/VCP. Moreover, we show that a p97-proteasome pathway model substrate (Ub(G76V)-GFP) is directly modified by K27-linked ubiquitylation, and that disabling the formation of K27-linked ubiquitin signals or blocking their decoding via overexpression of the K27 linkage-specific binder UCHL3 impedes Ub(G76V)-GFP turnover at the level of p97 function. Our findings suggest a critical role of K27-linked ubiquitylation in supporting cell fitness by facilitating p97-dependent processing of ubiquitylated nuclear proteins.
U2 - 10.15252/embj.2021110145
DO - 10.15252/embj.2021110145
M3 - Journal article
C2 - 35349166
VL - 41
JO - E M B O Journal
JF - E M B O Journal
SN - 0261-4189
IS - 9
M1 - e110145
ER -
ID: 303111768