Phospholipase C-related catalytically inactive protein participates in the autophagic elimination of Staphylococcus aureus infecting mouse embryonic fibroblasts.

Autophagy is an intrinsic host defense system that recognizes and eliminates invading bacterial pathogens. We have identified microtubule-associated protein 1 light chain 3 (LC3), a hallmark of autophagy, as a binding partner of phospholipase C-related catalytically inactive protein (PRIP) that was...

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Main Authors: Kae Harada-Hada, Kana Harada, Fuminori Kato, Junzo Hisatsune, Isei Tanida, Michinaga Ogawa, Satoshi Asano, Motoyuki Sugai, Masato Hirata, Takashi Kanematsu
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24865216/pdf/?tool=EBI
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author Kae Harada-Hada
Kana Harada
Fuminori Kato
Junzo Hisatsune
Isei Tanida
Michinaga Ogawa
Satoshi Asano
Motoyuki Sugai
Masato Hirata
Takashi Kanematsu
author_facet Kae Harada-Hada
Kana Harada
Fuminori Kato
Junzo Hisatsune
Isei Tanida
Michinaga Ogawa
Satoshi Asano
Motoyuki Sugai
Masato Hirata
Takashi Kanematsu
author_sort Kae Harada-Hada
collection DOAJ
description Autophagy is an intrinsic host defense system that recognizes and eliminates invading bacterial pathogens. We have identified microtubule-associated protein 1 light chain 3 (LC3), a hallmark of autophagy, as a binding partner of phospholipase C-related catalytically inactive protein (PRIP) that was originally identified as an inositol trisphosphate-binding protein. Here, we investigated the involvement of PRIP in the autophagic elimination of Staphylococcus aureus in infected mouse embryonic fibroblasts (MEFs). We observed significantly more LC3-positive autophagosome-like vacuoles enclosing an increased number of S. aureus cells in PRIP-deficient MEFs than control MEFs, 3 h and 4.5 h post infection, suggesting that S. aureus proliferates in LC3-positive autophagosome-like vacuoles in PRIP-deficient MEFs. We performed autophagic flux analysis using an mRFP-GFP-tagged LC3 plasmid and found that autophagosome maturation is significantly inhibited in PRIP-deficient MEFs. Furthermore, acidification of autophagosomes was significantly inhibited in PRIP-deficient MEFs compared to the wild-type MEFs, as determined by LysoTracker staining and time-lapse image analysis performed using mRFP-GFP-tagged LC3. Taken together, our data show that PRIP is required for the fusion of S. aureus-containing autophagosome-like vacuoles with lysosomes, indicating that PRIP is a novel modulator in the regulation of the innate immune system in non-professional phagocytic host cells.
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spelling doaj.art-0401bd79566a41b097b32e8da8151dac2022-12-21T16:58:12ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0195e9828510.1371/journal.pone.0098285Phospholipase C-related catalytically inactive protein participates in the autophagic elimination of Staphylococcus aureus infecting mouse embryonic fibroblasts.Kae Harada-HadaKana HaradaFuminori KatoJunzo HisatsuneIsei TanidaMichinaga OgawaSatoshi AsanoMotoyuki SugaiMasato HirataTakashi KanematsuAutophagy is an intrinsic host defense system that recognizes and eliminates invading bacterial pathogens. We have identified microtubule-associated protein 1 light chain 3 (LC3), a hallmark of autophagy, as a binding partner of phospholipase C-related catalytically inactive protein (PRIP) that was originally identified as an inositol trisphosphate-binding protein. Here, we investigated the involvement of PRIP in the autophagic elimination of Staphylococcus aureus in infected mouse embryonic fibroblasts (MEFs). We observed significantly more LC3-positive autophagosome-like vacuoles enclosing an increased number of S. aureus cells in PRIP-deficient MEFs than control MEFs, 3 h and 4.5 h post infection, suggesting that S. aureus proliferates in LC3-positive autophagosome-like vacuoles in PRIP-deficient MEFs. We performed autophagic flux analysis using an mRFP-GFP-tagged LC3 plasmid and found that autophagosome maturation is significantly inhibited in PRIP-deficient MEFs. Furthermore, acidification of autophagosomes was significantly inhibited in PRIP-deficient MEFs compared to the wild-type MEFs, as determined by LysoTracker staining and time-lapse image analysis performed using mRFP-GFP-tagged LC3. Taken together, our data show that PRIP is required for the fusion of S. aureus-containing autophagosome-like vacuoles with lysosomes, indicating that PRIP is a novel modulator in the regulation of the innate immune system in non-professional phagocytic host cells.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24865216/pdf/?tool=EBI
spellingShingle Kae Harada-Hada
Kana Harada
Fuminori Kato
Junzo Hisatsune
Isei Tanida
Michinaga Ogawa
Satoshi Asano
Motoyuki Sugai
Masato Hirata
Takashi Kanematsu
Phospholipase C-related catalytically inactive protein participates in the autophagic elimination of Staphylococcus aureus infecting mouse embryonic fibroblasts.
PLoS ONE
title Phospholipase C-related catalytically inactive protein participates in the autophagic elimination of Staphylococcus aureus infecting mouse embryonic fibroblasts.
title_full Phospholipase C-related catalytically inactive protein participates in the autophagic elimination of Staphylococcus aureus infecting mouse embryonic fibroblasts.
title_fullStr Phospholipase C-related catalytically inactive protein participates in the autophagic elimination of Staphylococcus aureus infecting mouse embryonic fibroblasts.
title_full_unstemmed Phospholipase C-related catalytically inactive protein participates in the autophagic elimination of Staphylococcus aureus infecting mouse embryonic fibroblasts.
title_short Phospholipase C-related catalytically inactive protein participates in the autophagic elimination of Staphylococcus aureus infecting mouse embryonic fibroblasts.
title_sort phospholipase c related catalytically inactive protein participates in the autophagic elimination of staphylococcus aureus infecting mouse embryonic fibroblasts
url https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24865216/pdf/?tool=EBI
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