Activity-independent targeting of mTOR to lysosomes in primary osteoclasts

Abstract Mammalian target of rapamycin (mTOR) is activated by numerous stimuli, including amino acids and growth factors. This kinase is part of the mTOR complex 1 (mTORC1) which regulates cell proliferation, differentiation, and autophagy. Active mTORC1 is located on lysosomes and has been reported...

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Autores principales:	Andrew Wang, Luciene R. Carraro-Lacroix, Celeste Owen, Bowen Gao, Paul N. Corey, Pascal Tyrrell, John H. Brumell, Irina Voronov
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/48c316c68a3b41edb20300c3142b91c4
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spelling	oai:doaj.org-article:48c316c68a3b41edb20300c3142b91c42021-12-02T11:52:19ZActivity-independent targeting of mTOR to lysosomes in primary osteoclasts10.1038/s41598-017-03494-22045-2322https://doaj.org/article/48c316c68a3b41edb20300c3142b91c42017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-03494-2https://doaj.org/toc/2045-2322Abstract Mammalian target of rapamycin (mTOR) is activated by numerous stimuli, including amino acids and growth factors. This kinase is part of the mTOR complex 1 (mTORC1) which regulates cell proliferation, differentiation, and autophagy. Active mTORC1 is located on lysosomes and has been reported to disassociate from the lysosomal surface in the absence of amino acids. Furthermore, mTORC1 activity has been linked to the vacuolar H+-ATPases (V-ATPases), the proton pumps responsible for lysosomal acidification; however, the exact role of the V-ATPases in mTORC1 signaling is not known. To elucidate the mechanisms involved in mTORC1 regulation by the V-ATPases, we used primary osteoclasts derived from mice carrying a point (R740S) mutation in the a3 subunit of the V-ATPase. In these cells, the mutant protein is expressed but the pump is not functional, resulting in higher lysosomal pH. By analyzing mTOR activation, mTOR/lysosome co-localization, and lysosomal positioning using confocal microscopy, fractionation, and ultrapure lysosomal purification methods, we demonstrate that in primary osteoclasts, mTOR is localized on the lysosomal surface even when mTOR activity is inhibited. Our findings reveal that mTOR targeting to the lysosome in osteoclasts is activity-independent, and that its disassociation from the lysosome during starvation is not universal.Andrew WangLuciene R. Carraro-LacroixCeleste OwenBowen GaoPaul N. CoreyPascal TyrrellJohn H. BrumellIrina VoronovNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Andrew Wang Luciene R. Carraro-Lacroix Celeste Owen Bowen Gao Paul N. Corey Pascal Tyrrell John H. Brumell Irina Voronov Activity-independent targeting of mTOR to lysosomes in primary osteoclasts
description	Abstract Mammalian target of rapamycin (mTOR) is activated by numerous stimuli, including amino acids and growth factors. This kinase is part of the mTOR complex 1 (mTORC1) which regulates cell proliferation, differentiation, and autophagy. Active mTORC1 is located on lysosomes and has been reported to disassociate from the lysosomal surface in the absence of amino acids. Furthermore, mTORC1 activity has been linked to the vacuolar H+-ATPases (V-ATPases), the proton pumps responsible for lysosomal acidification; however, the exact role of the V-ATPases in mTORC1 signaling is not known. To elucidate the mechanisms involved in mTORC1 regulation by the V-ATPases, we used primary osteoclasts derived from mice carrying a point (R740S) mutation in the a3 subunit of the V-ATPase. In these cells, the mutant protein is expressed but the pump is not functional, resulting in higher lysosomal pH. By analyzing mTOR activation, mTOR/lysosome co-localization, and lysosomal positioning using confocal microscopy, fractionation, and ultrapure lysosomal purification methods, we demonstrate that in primary osteoclasts, mTOR is localized on the lysosomal surface even when mTOR activity is inhibited. Our findings reveal that mTOR targeting to the lysosome in osteoclasts is activity-independent, and that its disassociation from the lysosome during starvation is not universal.
format	article
author	Andrew Wang Luciene R. Carraro-Lacroix Celeste Owen Bowen Gao Paul N. Corey Pascal Tyrrell John H. Brumell Irina Voronov
author_facet	Andrew Wang Luciene R. Carraro-Lacroix Celeste Owen Bowen Gao Paul N. Corey Pascal Tyrrell John H. Brumell Irina Voronov
author_sort	Andrew Wang
title	Activity-independent targeting of mTOR to lysosomes in primary osteoclasts
title_short	Activity-independent targeting of mTOR to lysosomes in primary osteoclasts
title_full	Activity-independent targeting of mTOR to lysosomes in primary osteoclasts
title_fullStr	Activity-independent targeting of mTOR to lysosomes in primary osteoclasts
title_full_unstemmed	Activity-independent targeting of mTOR to lysosomes in primary osteoclasts
title_sort	activity-independent targeting of mtor to lysosomes in primary osteoclasts
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/48c316c68a3b41edb20300c3142b91c4
work_keys_str_mv	AT andrewwang activityindependenttargetingofmtortolysosomesinprimaryosteoclasts AT lucienercarrarolacroix activityindependenttargetingofmtortolysosomesinprimaryosteoclasts AT celesteowen activityindependenttargetingofmtortolysosomesinprimaryosteoclasts AT bowengao activityindependenttargetingofmtortolysosomesinprimaryosteoclasts AT paulncorey activityindependenttargetingofmtortolysosomesinprimaryosteoclasts AT pascaltyrrell activityindependenttargetingofmtortolysosomesinprimaryosteoclasts AT johnhbrumell activityindependenttargetingofmtortolysosomesinprimaryosteoclasts AT irinavoronov activityindependenttargetingofmtortolysosomesinprimaryosteoclasts
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Activity-independent targeting of mTOR to lysosomes in primary osteoclasts

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