p62/SQSTM1 promotes rapid ubiquitin conjugation to target proteins after endosome rupture during xenophagy

Autophagy is a bulk degradation pathway, and selective autophagy to remove foreign entities is called xenophagy. The conjugation of ubiquitin to target pathogens is an important process in xenophagy but when and where this ubiquitination occurs remains unclear. Here, we analyzed the temporal sequenc...

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Main Authors: Megumi Tsuchiya, Hidesato Ogawa, Takako Koujin, Chie Mori, Hiroko Osakada, Shouhei Kobayashi, Yasushi Hiraoka, Tokuko Haraguchi
Format: Article
Language:English
Published: Wiley 2018-03-01
Series:FEBS Open Bio
Subjects:
Online Access:https://doi.org/10.1002/2211-5463.12385
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author Megumi Tsuchiya
Hidesato Ogawa
Takako Koujin
Chie Mori
Hiroko Osakada
Shouhei Kobayashi
Yasushi Hiraoka
Tokuko Haraguchi
author_facet Megumi Tsuchiya
Hidesato Ogawa
Takako Koujin
Chie Mori
Hiroko Osakada
Shouhei Kobayashi
Yasushi Hiraoka
Tokuko Haraguchi
author_sort Megumi Tsuchiya
collection DOAJ
description Autophagy is a bulk degradation pathway, and selective autophagy to remove foreign entities is called xenophagy. The conjugation of ubiquitin to target pathogens is an important process in xenophagy but when and where this ubiquitination occurs remains unclear. Here, we analyzed the temporal sequence and subcellular location of ubiquitination during xenophagy using time‐lapse observations, with polystyrene beads mimicking invading pathogens. Results revealed accumulation of a ubiquitination marker around the beads within 3 min after endosome rupture. Recruitment of ubiquitin to the beads was significantly delayed in p62‐knockout murine embryonic fibroblast cells, and this delay was rescued by ectopic p62 expression. Ectopic expression of a phosphorylation‐mimicking p62 mutated at serine residue 405 (equivalent to human serine residue 403) rescued this delay, but its unphosphorylated form did not. These results indicate that ubiquitination mainly occurs after endosome rupture and suggest that p62, specifically the phosphorylated form, promotes ubiquitin conjugation to target proteins in xenophagy.
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id doaj-art-7b7b1292dd3d4e959b42c96b1f95cae1
institution Kabale University
issn 2211-5463
language English
publishDate 2018-03-01
publisher Wiley
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series FEBS Open Bio
spelling doaj-art-7b7b1292dd3d4e959b42c96b1f95cae12024-12-26T06:31:12ZengWileyFEBS Open Bio2211-54632018-03-018347048010.1002/2211-5463.12385p62/SQSTM1 promotes rapid ubiquitin conjugation to target proteins after endosome rupture during xenophagyMegumi Tsuchiya0Hidesato Ogawa1Takako Koujin2Chie Mori3Hiroko Osakada4Shouhei Kobayashi5Yasushi Hiraoka6Tokuko Haraguchi7Graduate School of Frontier Biosciences Osaka University Suita JapanGraduate School of Frontier Biosciences Osaka University Suita JapanAdvanced ICT Research Institute Kobe National Institute of Information and Communications Technology Kobe JapanAdvanced ICT Research Institute Kobe National Institute of Information and Communications Technology Kobe JapanAdvanced ICT Research Institute Kobe National Institute of Information and Communications Technology Kobe JapanAdvanced ICT Research Institute Kobe National Institute of Information and Communications Technology Kobe JapanGraduate School of Frontier Biosciences Osaka University Suita JapanGraduate School of Frontier Biosciences Osaka University Suita JapanAutophagy is a bulk degradation pathway, and selective autophagy to remove foreign entities is called xenophagy. The conjugation of ubiquitin to target pathogens is an important process in xenophagy but when and where this ubiquitination occurs remains unclear. Here, we analyzed the temporal sequence and subcellular location of ubiquitination during xenophagy using time‐lapse observations, with polystyrene beads mimicking invading pathogens. Results revealed accumulation of a ubiquitination marker around the beads within 3 min after endosome rupture. Recruitment of ubiquitin to the beads was significantly delayed in p62‐knockout murine embryonic fibroblast cells, and this delay was rescued by ectopic p62 expression. Ectopic expression of a phosphorylation‐mimicking p62 mutated at serine residue 405 (equivalent to human serine residue 403) rescued this delay, but its unphosphorylated form did not. These results indicate that ubiquitination mainly occurs after endosome rupture and suggest that p62, specifically the phosphorylated form, promotes ubiquitin conjugation to target proteins in xenophagy.https://doi.org/10.1002/2211-5463.12385artificial beadsautophagybacterial infectionLC3p62ubiquitination
spellingShingle Megumi Tsuchiya
Hidesato Ogawa
Takako Koujin
Chie Mori
Hiroko Osakada
Shouhei Kobayashi
Yasushi Hiraoka
Tokuko Haraguchi
p62/SQSTM1 promotes rapid ubiquitin conjugation to target proteins after endosome rupture during xenophagy
FEBS Open Bio
artificial beads
autophagy
bacterial infection
LC3
p62
ubiquitination
title p62/SQSTM1 promotes rapid ubiquitin conjugation to target proteins after endosome rupture during xenophagy
title_full p62/SQSTM1 promotes rapid ubiquitin conjugation to target proteins after endosome rupture during xenophagy
title_fullStr p62/SQSTM1 promotes rapid ubiquitin conjugation to target proteins after endosome rupture during xenophagy
title_full_unstemmed p62/SQSTM1 promotes rapid ubiquitin conjugation to target proteins after endosome rupture during xenophagy
title_short p62/SQSTM1 promotes rapid ubiquitin conjugation to target proteins after endosome rupture during xenophagy
title_sort p62 sqstm1 promotes rapid ubiquitin conjugation to target proteins after endosome rupture during xenophagy
topic artificial beads
autophagy
bacterial infection
LC3
p62
ubiquitination
url https://doi.org/10.1002/2211-5463.12385
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