Unknown

Dataset Information

0

Novel polyubiquitin imaging system, PolyUb-FC, reveals that K33-linked polyubiquitin is recruited by SQSTM1/p62.


ABSTRACT: Ubiquitin chains are formed with 8 structurally and functionally distinct polymers. However, the functions of each polyubiquitin remain poorly understood. We developed a polyubiquitin-mediated fluorescence complementation (PolyUb-FC) assay using Kusabira Green (KG) as a split fluorescent protein. The PolyUb-FC assay has the advantage that monoubiquitination is nonfluorescent and chain-specific polyubiquitination can be directly visualized in living cells without using antibodies. We applied the PolyUb-FC assay to examine K33-linked polyubiquitin. We demonstrated that SQSTM1/p62 puncta colocalized with K33-linked polyubiquitin and this interaction was modulated by the ZRANB1/TRABID-K29 and -K33 linkage-specific deubiquitinase (DUB). We further showed that the colocalization of K33-linked polyubiquitin and MAP1LC3/LC3 (microtubule associated protein 1 light chain 3) puncta was impaired by SQSTM1/p62 deficiency. Taken together, these findings provide novel insights into how atypical polyubiquitin is recruited by SQSTM1/p62. Finally, we developed an inducible-PolyUb-FC system for visualizing chain-specific polyubiquitin. The PolyUb-FC will be a useful tool for analyzing the dynamics of atypical polyubiquitin chain generation.

SUBMITTER: Nibe Y 

PROVIDER: S-EPMC5902196 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

altmetric image

Publications


Ubiquitin chains are formed with 8 structurally and functionally distinct polymers. However, the functions of each polyubiquitin remain poorly understood. We developed a polyubiquitin-mediated fluorescence complementation (PolyUb-FC) assay using Kusabira Green (KG) as a split fluorescent protein. The PolyUb-FC assay has the advantage that monoubiquitination is nonfluorescent and chain-specific polyubiquitination can be directly visualized in living cells without using antibodies. We applied the  ...[more]

Similar Datasets

| S-EPMC5640208 | biostudies-literature
| S-SCDT-EMBOR-2020-51136V1 | biostudies-other
| S-EPMC2581430 | biostudies-literature
2018-03-31 | E-MTAB-4894 | biostudies-arrayexpress
| S-EPMC4390085 | biostudies-literature
| S-EPMC7144840 | biostudies-literature
| S-EPMC6197226 | biostudies-literature
| S-EPMC3782476 | biostudies-literature
| S-EPMC4627259 | biostudies-literature
2022-07-21 | GSE208588 | GEO