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PACRG and FAP20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility
We previously demonstrated that PACRG plays a role in regulating dynein-driven microtubule sliding in motile cilia. To expand our understanding of the role of PACRG in ciliary assembly and motility, we used a combination of functional and structural studies, including newly identified Chlamydomonas...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6727744/ https://www.ncbi.nlm.nih.gov/pubmed/31116684 http://dx.doi.org/10.1091/mbc.E19-01-0063 |
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author | Dymek, Erin E. Lin, Jianfeng Fu, Gang Porter, Mary E. Nicastro, Daniela Smith, Elizabeth F. |
author_facet | Dymek, Erin E. Lin, Jianfeng Fu, Gang Porter, Mary E. Nicastro, Daniela Smith, Elizabeth F. |
author_sort | Dymek, Erin E. |
collection | PubMed |
description | We previously demonstrated that PACRG plays a role in regulating dynein-driven microtubule sliding in motile cilia. To expand our understanding of the role of PACRG in ciliary assembly and motility, we used a combination of functional and structural studies, including newly identified Chlamydomonas pacrg mutants. Using cryo-electron tomography we show that PACRG and FAP20 form the inner junction between the A- and B-tubule along the length of all nine ciliary doublet microtubules. The lack of PACRG and FAP20 also results in reduced assembly of inner-arm dynein IDA b and the beak-MIP structures. In addition, our functional studies reveal that loss of PACRG and/or FAP20 causes severe cell motility defects and reduced in vitro microtubule sliding velocities. Interestingly, the addition of exogenous PACRG and/or FAP20 protein to isolated mutant axonemes restores microtubule sliding velocities, but not ciliary beating. Taken together, these studies show that PACRG and FAP20 comprise the inner junction bridge that serves as a hub for both directly modulating dynein-driven microtubule sliding, as well as for the assembly of additional ciliary components that play essential roles in generating coordinated ciliary beating. |
format | Online Article Text |
id | pubmed-6727744 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-67277442019-09-30 PACRG and FAP20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility Dymek, Erin E. Lin, Jianfeng Fu, Gang Porter, Mary E. Nicastro, Daniela Smith, Elizabeth F. Mol Biol Cell Articles We previously demonstrated that PACRG plays a role in regulating dynein-driven microtubule sliding in motile cilia. To expand our understanding of the role of PACRG in ciliary assembly and motility, we used a combination of functional and structural studies, including newly identified Chlamydomonas pacrg mutants. Using cryo-electron tomography we show that PACRG and FAP20 form the inner junction between the A- and B-tubule along the length of all nine ciliary doublet microtubules. The lack of PACRG and FAP20 also results in reduced assembly of inner-arm dynein IDA b and the beak-MIP structures. In addition, our functional studies reveal that loss of PACRG and/or FAP20 causes severe cell motility defects and reduced in vitro microtubule sliding velocities. Interestingly, the addition of exogenous PACRG and/or FAP20 protein to isolated mutant axonemes restores microtubule sliding velocities, but not ciliary beating. Taken together, these studies show that PACRG and FAP20 comprise the inner junction bridge that serves as a hub for both directly modulating dynein-driven microtubule sliding, as well as for the assembly of additional ciliary components that play essential roles in generating coordinated ciliary beating. The American Society for Cell Biology 2019-07-15 /pmc/articles/PMC6727744/ /pubmed/31116684 http://dx.doi.org/10.1091/mbc.E19-01-0063 Text en © 2019 Dymek et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0 This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License. |
spellingShingle | Articles Dymek, Erin E. Lin, Jianfeng Fu, Gang Porter, Mary E. Nicastro, Daniela Smith, Elizabeth F. PACRG and FAP20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility |
title | PACRG and FAP20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility |
title_full | PACRG and FAP20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility |
title_fullStr | PACRG and FAP20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility |
title_full_unstemmed | PACRG and FAP20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility |
title_short | PACRG and FAP20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility |
title_sort | pacrg and fap20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6727744/ https://www.ncbi.nlm.nih.gov/pubmed/31116684 http://dx.doi.org/10.1091/mbc.E19-01-0063 |
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