Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis

A Disintegrin and Metalloproteinase with ThromboSpondin motif (ADAMTS) 5 functions as an anti-angiogenic and anti-cancer protein independent of its metalloproteinase activity. Both full-length ADAMTS5 and TS5-p45, the autocatalytically cleaved C-terminal 45 kDa truncate of ADAMTS5, inhibits angiogen...

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Main Authors: Kirman, Dogan Can, Renganathan, Bhuvanasundar, Chui, Wai Kit, Chen, Ming Wei, Kaya, Neslihan Arife, Ge, Ruowen
Other Authors: School of Biological Sciences
Format: Article
Language:English
Published: 2022
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Online Access:https://hdl.handle.net/10356/163244
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Institution: Nanyang Technological University
Language: English
id sg-ntu-dr.10356-163244
record_format dspace
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Science::Biological sciences
Phosphoprotein
Antiangiogenic activity
spellingShingle Science::Biological sciences
Phosphoprotein
Antiangiogenic activity
Kirman, Dogan Can
Renganathan, Bhuvanasundar
Chui, Wai Kit
Chen, Ming Wei
Kaya, Neslihan Arife
Ge, Ruowen
Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis
description A Disintegrin and Metalloproteinase with ThromboSpondin motif (ADAMTS) 5 functions as an anti-angiogenic and anti-cancer protein independent of its metalloproteinase activity. Both full-length ADAMTS5 and TS5-p45, the autocatalytically cleaved C-terminal 45 kDa truncate of ADAMTS5, inhibits angiogenesis, and induces endothelial cell (EC) apoptosis. However, how ADAMTS5 triggers EC apoptosis remains unclear. This work shows that caspase-8 (Cas-8) and caspase-9 (Cas-9) are involved in TS5-p45-induced EC apoptosis. We identify cell surface nucleolin (NCL) as a novel high-affinity receptor for TS5-p45 in ECs, mediating TS5-p45's cell surface binding and pro-apoptotic function. We show that the central RNA-binding domain (RBD) of NCL is essential and sufficient for its binding to TS5-p45. Upon interacting with EC surface NCL, TS5-p45 is internalized through clathrin- and caveolin-dependent endocytosis and trafficked to the nucleus via late endosomes (LEs). We demonstrate that the nuclear trafficking of TS5-p45 is important for its pro-apoptotic activity as disruption of LE membrane integrity with an endosomolytic peptide suppressed both nuclear trafficking and pro-apoptotic activity of TS5-p45. Through cell surface biotinylation, we revealed that cell surface NCL shuttles extracellular TS5-p45 to the nucleus to mediate apoptosis. Furthermore, blocking the importin α1/ß1 receptor hindered the nuclear trafficking of TS5-p45, suggesting the involvement of the nuclear importing machinery for this nuclear translocation. RNA-seq identified many apoptosis-related genes that are differentially expressed at least two-fold in TS5-p45-treated ECs, with 10 of them qRT-PCR-validated and at least 5 of these genes potentially contributing to TS5-p45-NCL-induced apoptosis. Altogether, our work identifies NCL as a novel cell surface receptor for ADAMTS5 and demonstrates the critical role of NCL-mediated internalization and nuclear trafficking for ADAMTS5-induced EC apoptosis. These findings reveal novel mechanistic insights of the secreted metalloproteinase ADAMTS5 in angiogenesis inhibition.
author2 School of Biological Sciences
author_facet School of Biological Sciences
Kirman, Dogan Can
Renganathan, Bhuvanasundar
Chui, Wai Kit
Chen, Ming Wei
Kaya, Neslihan Arife
Ge, Ruowen
format Article
author Kirman, Dogan Can
Renganathan, Bhuvanasundar
Chui, Wai Kit
Chen, Ming Wei
Kaya, Neslihan Arife
Ge, Ruowen
author_sort Kirman, Dogan Can
title Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis
title_short Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis
title_full Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis
title_fullStr Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis
title_full_unstemmed Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis
title_sort cell surface nucleolin is a novel adamts5 receptor mediating endothelial cell apoptosis
publishDate 2022
url https://hdl.handle.net/10356/163244
_version_ 1759856598480584704
spelling sg-ntu-dr.10356-1632442023-02-28T17:12:30Z Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis Kirman, Dogan Can Renganathan, Bhuvanasundar Chui, Wai Kit Chen, Ming Wei Kaya, Neslihan Arife Ge, Ruowen School of Biological Sciences Genome Institute of Singapore, A*STAR Science::Biological sciences Phosphoprotein Antiangiogenic activity A Disintegrin and Metalloproteinase with ThromboSpondin motif (ADAMTS) 5 functions as an anti-angiogenic and anti-cancer protein independent of its metalloproteinase activity. Both full-length ADAMTS5 and TS5-p45, the autocatalytically cleaved C-terminal 45 kDa truncate of ADAMTS5, inhibits angiogenesis, and induces endothelial cell (EC) apoptosis. However, how ADAMTS5 triggers EC apoptosis remains unclear. This work shows that caspase-8 (Cas-8) and caspase-9 (Cas-9) are involved in TS5-p45-induced EC apoptosis. We identify cell surface nucleolin (NCL) as a novel high-affinity receptor for TS5-p45 in ECs, mediating TS5-p45's cell surface binding and pro-apoptotic function. We show that the central RNA-binding domain (RBD) of NCL is essential and sufficient for its binding to TS5-p45. Upon interacting with EC surface NCL, TS5-p45 is internalized through clathrin- and caveolin-dependent endocytosis and trafficked to the nucleus via late endosomes (LEs). We demonstrate that the nuclear trafficking of TS5-p45 is important for its pro-apoptotic activity as disruption of LE membrane integrity with an endosomolytic peptide suppressed both nuclear trafficking and pro-apoptotic activity of TS5-p45. Through cell surface biotinylation, we revealed that cell surface NCL shuttles extracellular TS5-p45 to the nucleus to mediate apoptosis. Furthermore, blocking the importin α1/ß1 receptor hindered the nuclear trafficking of TS5-p45, suggesting the involvement of the nuclear importing machinery for this nuclear translocation. RNA-seq identified many apoptosis-related genes that are differentially expressed at least two-fold in TS5-p45-treated ECs, with 10 of them qRT-PCR-validated and at least 5 of these genes potentially contributing to TS5-p45-NCL-induced apoptosis. Altogether, our work identifies NCL as a novel cell surface receptor for ADAMTS5 and demonstrates the critical role of NCL-mediated internalization and nuclear trafficking for ADAMTS5-induced EC apoptosis. These findings reveal novel mechanistic insights of the secreted metalloproteinase ADAMTS5 in angiogenesis inhibition. Published version This work was supported by a grant awarded from the Singapore Ministry of Education to Ruowen Ge (MOE2014-T2-2-150). DCK was supported by the Singapore International Graduate Award from the Agency for Science, Technology & Research and the National University of Singapore. 2022-11-29T06:34:55Z 2022-11-29T06:34:55Z 2022 Journal Article Kirman, D. C., Renganathan, B., Chui, W. K., Chen, M. W., Kaya, N. A. & Ge, R. (2022). Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis. Cell Death and Disease, 13(2), 172-. https://dx.doi.org/10.1038/s41419-022-04618-x 2041-4889 https://hdl.handle.net/10356/163244 10.1038/s41419-022-04618-x 35197459 2-s2.0-85125156475 2 13 172 en Cell Death and Disease © The Author(s) 2022. Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. application/pdf