Giantin is required for intracellular N-terminal processing of type I procollagen

Nicola L Stevenson, Dylan J M Bergen, Esther Prada-Sanchez, Christina Hammond, David J Stephens

Research output: Contribution to journalArticle (Academic Journal)peer-review

3 Citations (Scopus)
183 Downloads (Pure)

Abstract

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Knockout of the golgin giantin leads to skeletal and craniofacial defects driven by poorly studied changes in glycosylation and extracellular matrix deposition. Here, we sought to determine how giantin impacts the production of healthy bone tissue by focusing on the main protein component of the osteoid, type I collagen. Giantin mutant zebrafish accumulate multiple spontaneous fractures in their caudal fin, suggesting their bones may be more brittle. Inducing new experimental fractures revealed defects in the mineralization of newly deposited collagen as well as diminished procollagen reporter expression in mutant fish. Analysis of a human giantin knockout cell line expressing a GFP-tagged procollagen showed that procollagen trafficking is independent of giantin. However, our data show that intracellular N-propeptide processing of pro-α1(I) is defective in the absence of giantin. These data demonstrate a conserved role for giantin in collagen biosynthesis and extracellular matrix assembly. Our work also provides evidence of a giantin-dependent pathway for intracellular procollagen processing.
Original languageEnglish
Article numbere202005166
Number of pages18
JournalJournal of Cell Biology
Volume220
Issue number6
DOIs
Publication statusPublished - 7 Jun 2021

Bibliographical note

Funding Information:
We thank Franck Perez and Gaelle Boncompain (Institut Curie, France) for sharing the RUSH system with us, Helen Dawe (University of Exeter, UK) and Stuart Haslam (Imperial College London, UK) for their technical help and advice, Andrew Her-man (University of Bristol flow cytometry facility) and Kate Heesom (University of Bristol proteomics facility) for their work on this project, Lucy McGowan for help with zebrafish experiments, and Alain Colige (University of Liege, Liege, Belgium) for his help and advice and for providing an ADAMTS2 construct. We also thank the Medical Research Council and Wolfson Foundation for establishing the Wolfson Bioimaging Facility. This work was funded by the UK Research and Innovation? Medical Research Council (MR/P000177/1), Versus Arthritis (21937 and 22044), and UK Research and Innovation? Biotechnology and Biological Sciences Research Council (BB/ T001984/1). Confocal microscopy was also supported by UK Research and Innovation?Biotechnology and Biological Sciences Research Council (BB/L014181/1) and electron microscopy by the Wellcome Trust (110126/Z/15/Z).

Funding Information:
We thank Franck Perez and Gaelle Boncompain (Institut Curie, France) for sharing the RUSH system with us, Helen Dawe (University of Exeter, UK) and Stuart Haslam (Imperial College London, UK) for their technical help and advice, Andrew Herman (University of Bristol flow cytometry facility) and Kate Heesom (University of Bristol proteomics facility) for their work on this project, Lucy McGowan for help with zebrafish experiments, and Alain Colige (University of Liege, Liege, Belgium) for his help and advice and for providing an ADAMTS2 construct. We also thank the Medical Research Council and Wolfson Foundation for establishing the Wolfson Bioimaging Facility.

Funding Information:
This work was funded by the UK Research and Innovation– Medical Research Council (MR/P000177/1), Versus Arthritis (21937 and 22044), and UK Research and Innovation– Biotechnology and Biological Sciences Research Council (BB/ T001984/1). Confocal microscopy was also supported by UK Research and Innovation–Biotechnology and Biological Sciences Research Council (BB/L014181/1) and electron microscopy by the Wellcome Trust (110126/Z/15/Z). The authors declare no competing financial interests.

Publisher Copyright:
© 2021 Stevenson et al.

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