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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...

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Autores principales: Dymek, Erin E., Lin, Jianfeng, Fu, Gang, Porter, Mary E., Nicastro, Daniela, Smith, Elizabeth F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2019
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.
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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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