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TitlePACRG and FAP20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility.
Journal, issue, pagesMol Biol Cell, Vol. 30, Issue 15, Page 1805-1816, Year 2019
Publish dateJul 15, 2019
AuthorsErin E Dymek / Jianfeng Lin / Gang Fu / Mary E Porter / Daniela Nicastro / Elizabeth F Smith /
PubMed AbstractWe 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 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 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 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.
External linksMol Biol Cell / PubMed:31116684 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution23.0 - 27.0 Å
Structure data

EMDB-0628:
The axonemal doublet microtubule focusing on the inner junction region extracted from the cryo-electron tomography and subtomographic average of isolated Chlamydomonas wild type cilia
Method: EM (subtomogram averaging) / Resolution: 23.0 Å

EMDB-0629:
The axonemal doublet microtubule focusing on the inner junction region extracted from the cryo-electron tomography and subtomographic average of isolated Chlamydomonas pacrg mutant cilia
Method: EM (subtomogram averaging) / Resolution: 27.0 Å

EMDB-0630:
The axonemal doublet microtubule focusing on the inner junction region extracted from the cryo-electron tomography and subtomographic average of isolated Chlamydomonas pacrg; fap20 double mutant cilia
Method: EM (subtomogram averaging) / Resolution: 24.0 Å

Source
  • Chlamydomonas reinhardtii (plant)

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