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CCP5 and CCP6 retain CP110 and negatively regulate ciliogenesis
BACKGROUND: The axonemal microtubules of primary cilium undergo a conserved protein posttranslational modification (PTM) — polyglutamylation. This reversible procedure is processed by tubulin tyrosine ligase-like polyglutamylases to form secondary polyglutamate side chains, which are metabolized by...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10210458/ https://www.ncbi.nlm.nih.gov/pubmed/37226238 http://dx.doi.org/10.1186/s12915-023-01622-1 |
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author | Wang, Yujuan Zhang, Yuan Guo, Xinyu Zheng, Yiqiang Zhang, Xinjie Feng, Shanshan Wu, Hui-Yuan |
author_facet | Wang, Yujuan Zhang, Yuan Guo, Xinyu Zheng, Yiqiang Zhang, Xinjie Feng, Shanshan Wu, Hui-Yuan |
author_sort | Wang, Yujuan |
collection | PubMed |
description | BACKGROUND: The axonemal microtubules of primary cilium undergo a conserved protein posttranslational modification (PTM) — polyglutamylation. This reversible procedure is processed by tubulin tyrosine ligase-like polyglutamylases to form secondary polyglutamate side chains, which are metabolized by the 6-member cytosolic carboxypeptidase (CCP) family. Although polyglutamylation modifying enzymes have been linked to ciliary architecture and motility, it was unknown whether they also play a role in ciliogenesis. RESULTS: In this study, we found that CCP5 expression is transiently downregulated upon the initiation of ciliogenesis, but recovered after cilia are formed. Overexpression of CCP5 inhibited ciliogenesis, suggesting that a transient downregulation of CCP5 expression is required for ciliation initiation. Interestingly, the inhibitory effect of CCP5 on ciliogenesis does not rely on its enzyme activity. Among other 3 CCP members tested, only CCP6 can similarly suppress ciliogenesis. Using CoIP-MS analysis, we identified a protein that potentially interacts with CCP — CP110, a known negative regulator of ciliogenesis, whose degradation at the distal end of mother centriole permits cilia assembly. We found that both CCP5 and CCP6 can modulate CP110 level. Particularly, CCP5 interacts with CP110 through its N-terminus. Loss of CCP5 or CCP6 led to the disappearance of CP110 at the mother centriole and abnormally increased ciliation in cycling RPE-1 cells. Co-depletion of CCP5 and CCP6 synergized this abnormal ciliation, suggesting their partially overlapped function in suppressing cilia formation in cycling cells. In contrast, co-depletion of the two enzymes did not further increase the length of cilia, although CCP5 and CCP6 differentially regulate polyglutamate side-chain length of ciliary axoneme and both contribute to limiting cilia length, suggesting that they may share a common pathway in cilia length control. Through inducing the overexpression of CCP5 or CCP6 at different stages of ciliogenesis, we further demonstrated that CCP5 or CCP6 inhibited cilia formation before ciliogenesis, while shortened the length of cilia after cilia formation. CONCLUSION: These findings reveal the dual role of CCP5 and CCP6. In addition to regulating cilia length, they also retain CP110 level to suppress cilia formation in cycling cells, pointing to a novel regulatory mechanism for ciliogenesis mediated by demodifying enzymes of a conserved ciliary PTM, polyglutamylation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12915-023-01622-1. |
format | Online Article Text |
id | pubmed-10210458 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-102104582023-05-26 CCP5 and CCP6 retain CP110 and negatively regulate ciliogenesis Wang, Yujuan Zhang, Yuan Guo, Xinyu Zheng, Yiqiang Zhang, Xinjie Feng, Shanshan Wu, Hui-Yuan BMC Biol Research Article BACKGROUND: The axonemal microtubules of primary cilium undergo a conserved protein posttranslational modification (PTM) — polyglutamylation. This reversible procedure is processed by tubulin tyrosine ligase-like polyglutamylases to form secondary polyglutamate side chains, which are metabolized by the 6-member cytosolic carboxypeptidase (CCP) family. Although polyglutamylation modifying enzymes have been linked to ciliary architecture and motility, it was unknown whether they also play a role in ciliogenesis. RESULTS: In this study, we found that CCP5 expression is transiently downregulated upon the initiation of ciliogenesis, but recovered after cilia are formed. Overexpression of CCP5 inhibited ciliogenesis, suggesting that a transient downregulation of CCP5 expression is required for ciliation initiation. Interestingly, the inhibitory effect of CCP5 on ciliogenesis does not rely on its enzyme activity. Among other 3 CCP members tested, only CCP6 can similarly suppress ciliogenesis. Using CoIP-MS analysis, we identified a protein that potentially interacts with CCP — CP110, a known negative regulator of ciliogenesis, whose degradation at the distal end of mother centriole permits cilia assembly. We found that both CCP5 and CCP6 can modulate CP110 level. Particularly, CCP5 interacts with CP110 through its N-terminus. Loss of CCP5 or CCP6 led to the disappearance of CP110 at the mother centriole and abnormally increased ciliation in cycling RPE-1 cells. Co-depletion of CCP5 and CCP6 synergized this abnormal ciliation, suggesting their partially overlapped function in suppressing cilia formation in cycling cells. In contrast, co-depletion of the two enzymes did not further increase the length of cilia, although CCP5 and CCP6 differentially regulate polyglutamate side-chain length of ciliary axoneme and both contribute to limiting cilia length, suggesting that they may share a common pathway in cilia length control. Through inducing the overexpression of CCP5 or CCP6 at different stages of ciliogenesis, we further demonstrated that CCP5 or CCP6 inhibited cilia formation before ciliogenesis, while shortened the length of cilia after cilia formation. CONCLUSION: These findings reveal the dual role of CCP5 and CCP6. In addition to regulating cilia length, they also retain CP110 level to suppress cilia formation in cycling cells, pointing to a novel regulatory mechanism for ciliogenesis mediated by demodifying enzymes of a conserved ciliary PTM, polyglutamylation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12915-023-01622-1. BioMed Central 2023-05-24 /pmc/articles/PMC10210458/ /pubmed/37226238 http://dx.doi.org/10.1186/s12915-023-01622-1 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Article Wang, Yujuan Zhang, Yuan Guo, Xinyu Zheng, Yiqiang Zhang, Xinjie Feng, Shanshan Wu, Hui-Yuan CCP5 and CCP6 retain CP110 and negatively regulate ciliogenesis |
title | CCP5 and CCP6 retain CP110 and negatively regulate ciliogenesis |
title_full | CCP5 and CCP6 retain CP110 and negatively regulate ciliogenesis |
title_fullStr | CCP5 and CCP6 retain CP110 and negatively regulate ciliogenesis |
title_full_unstemmed | CCP5 and CCP6 retain CP110 and negatively regulate ciliogenesis |
title_short | CCP5 and CCP6 retain CP110 and negatively regulate ciliogenesis |
title_sort | ccp5 and ccp6 retain cp110 and negatively regulate ciliogenesis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10210458/ https://www.ncbi.nlm.nih.gov/pubmed/37226238 http://dx.doi.org/10.1186/s12915-023-01622-1 |
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