Scaffold subunits support associated subunit assembly in the Chlamydomonas ciliary nexin-dynein regulatory complex

Long Gui, Kangkang Song, Douglas Tritschler, Raqual Bower, Si Yan, Aguang Dai, Katherine Augspurger, Jason Sakizadeh, Magdalena Grzemska, Thomas Ni, Mary E. Porter, Daniela Nicastro

Research output: Contribution to journalArticle

Abstract

The nexin-dynein regulatory complex (N-DRC) in motile cilia and flagella functions as a linker between neighboring doublet microtubules, acts to stabilize the axonemal core structure, and serves as a central hub for the regulation of ciliary motility. Although the N-DRC has been studied extensively using genetic, biochemical, and structural approaches, the precise arrangement of the 11 (or more) N-DRC subunits remains unknown. Here, using cryo-electron tomography, we have compared the structure of Chlamydomonas wild-type flagella to that of strains with specific DRC subunit deletions or rescued strains with tagged DRC subunits. Our results show that DRC7 is a central linker subunit that helps connect the N-DRC to the outer dynein arms. DRC11 is required for the assembly of DRC8, and DRC8/11 form a subcomplex in the proximal lobe of the linker domain that is required to form stable contacts to the neighboring B-tubule. Gold labeling of tagged subunits determines the precise locations of the previously ambiguous N terminus of DRC4 and C terminus of DRC5. DRC4 is now shown to contribute to the core scaffold of the N-DRC. Our results reveal the overall architecture of N-DRC, with the 3 subunits DRC1/2/4 forming a core complex that serves as the scaffold for the assembly of the "functional subunits," namely DRC3/5-8/11. These findings shed light on N-DRC assembly and its role in regulating flagellar beating.

Original languageEnglish (US)
Pages (from-to)23152-23162
Number of pages11
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number46
DOIs
StatePublished - Nov 12 2019

Fingerprint

Chlamydomonas
Dyneins
Flagella
Electron Microscope Tomography
Cilia
Microtubules
Gold
Molecular Biology

Keywords

  • Axoneme
  • Cilia
  • Cryo-electron tomography
  • Flagella
  • N-DRC

Cite this

Scaffold subunits support associated subunit assembly in the Chlamydomonas ciliary nexin-dynein regulatory complex. / Gui, Long; Song, Kangkang; Tritschler, Douglas; Bower, Raqual; Yan, Si; Dai, Aguang; Augspurger, Katherine; Sakizadeh, Jason; Grzemska, Magdalena; Ni, Thomas; Porter, Mary E.; Nicastro, Daniela.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 46, 12.11.2019, p. 23152-23162.

Research output: Contribution to journalArticle

Gui, L, Song, K, Tritschler, D, Bower, R, Yan, S, Dai, A, Augspurger, K, Sakizadeh, J, Grzemska, M, Ni, T, Porter, ME & Nicastro, D 2019, 'Scaffold subunits support associated subunit assembly in the Chlamydomonas ciliary nexin-dynein regulatory complex', Proceedings of the National Academy of Sciences of the United States of America, vol. 116, no. 46, pp. 23152-23162. https://doi.org/10.1073/pnas.1910960116
Gui, Long ; Song, Kangkang ; Tritschler, Douglas ; Bower, Raqual ; Yan, Si ; Dai, Aguang ; Augspurger, Katherine ; Sakizadeh, Jason ; Grzemska, Magdalena ; Ni, Thomas ; Porter, Mary E. ; Nicastro, Daniela. / Scaffold subunits support associated subunit assembly in the Chlamydomonas ciliary nexin-dynein regulatory complex. In: Proceedings of the National Academy of Sciences of the United States of America. 2019 ; Vol. 116, No. 46. pp. 23152-23162.
@article{86a3584f8ed74b9aa1d07394dd18395c,
title = "Scaffold subunits support associated subunit assembly in the Chlamydomonas ciliary nexin-dynein regulatory complex",
abstract = "The nexin-dynein regulatory complex (N-DRC) in motile cilia and flagella functions as a linker between neighboring doublet microtubules, acts to stabilize the axonemal core structure, and serves as a central hub for the regulation of ciliary motility. Although the N-DRC has been studied extensively using genetic, biochemical, and structural approaches, the precise arrangement of the 11 (or more) N-DRC subunits remains unknown. Here, using cryo-electron tomography, we have compared the structure of Chlamydomonas wild-type flagella to that of strains with specific DRC subunit deletions or rescued strains with tagged DRC subunits. Our results show that DRC7 is a central linker subunit that helps connect the N-DRC to the outer dynein arms. DRC11 is required for the assembly of DRC8, and DRC8/11 form a subcomplex in the proximal lobe of the linker domain that is required to form stable contacts to the neighboring B-tubule. Gold labeling of tagged subunits determines the precise locations of the previously ambiguous N terminus of DRC4 and C terminus of DRC5. DRC4 is now shown to contribute to the core scaffold of the N-DRC. Our results reveal the overall architecture of N-DRC, with the 3 subunits DRC1/2/4 forming a core complex that serves as the scaffold for the assembly of the {"}functional subunits,{"} namely DRC3/5-8/11. These findings shed light on N-DRC assembly and its role in regulating flagellar beating.",
keywords = "Axoneme, Cilia, Cryo-electron tomography, Flagella, N-DRC",
author = "Long Gui and Kangkang Song and Douglas Tritschler and Raqual Bower and Si Yan and Aguang Dai and Katherine Augspurger and Jason Sakizadeh and Magdalena Grzemska and Thomas Ni and Porter, {Mary E.} and Daniela Nicastro",
year = "2019",
month = "11",
day = "12",
doi = "10.1073/pnas.1910960116",
language = "English (US)",
volume = "116",
pages = "23152--23162",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "46",

}

TY - JOUR

T1 - Scaffold subunits support associated subunit assembly in the Chlamydomonas ciliary nexin-dynein regulatory complex

AU - Gui, Long

AU - Song, Kangkang

AU - Tritschler, Douglas

AU - Bower, Raqual

AU - Yan, Si

AU - Dai, Aguang

AU - Augspurger, Katherine

AU - Sakizadeh, Jason

AU - Grzemska, Magdalena

AU - Ni, Thomas

AU - Porter, Mary E.

AU - Nicastro, Daniela

PY - 2019/11/12

Y1 - 2019/11/12

N2 - The nexin-dynein regulatory complex (N-DRC) in motile cilia and flagella functions as a linker between neighboring doublet microtubules, acts to stabilize the axonemal core structure, and serves as a central hub for the regulation of ciliary motility. Although the N-DRC has been studied extensively using genetic, biochemical, and structural approaches, the precise arrangement of the 11 (or more) N-DRC subunits remains unknown. Here, using cryo-electron tomography, we have compared the structure of Chlamydomonas wild-type flagella to that of strains with specific DRC subunit deletions or rescued strains with tagged DRC subunits. Our results show that DRC7 is a central linker subunit that helps connect the N-DRC to the outer dynein arms. DRC11 is required for the assembly of DRC8, and DRC8/11 form a subcomplex in the proximal lobe of the linker domain that is required to form stable contacts to the neighboring B-tubule. Gold labeling of tagged subunits determines the precise locations of the previously ambiguous N terminus of DRC4 and C terminus of DRC5. DRC4 is now shown to contribute to the core scaffold of the N-DRC. Our results reveal the overall architecture of N-DRC, with the 3 subunits DRC1/2/4 forming a core complex that serves as the scaffold for the assembly of the "functional subunits," namely DRC3/5-8/11. These findings shed light on N-DRC assembly and its role in regulating flagellar beating.

AB - The nexin-dynein regulatory complex (N-DRC) in motile cilia and flagella functions as a linker between neighboring doublet microtubules, acts to stabilize the axonemal core structure, and serves as a central hub for the regulation of ciliary motility. Although the N-DRC has been studied extensively using genetic, biochemical, and structural approaches, the precise arrangement of the 11 (or more) N-DRC subunits remains unknown. Here, using cryo-electron tomography, we have compared the structure of Chlamydomonas wild-type flagella to that of strains with specific DRC subunit deletions or rescued strains with tagged DRC subunits. Our results show that DRC7 is a central linker subunit that helps connect the N-DRC to the outer dynein arms. DRC11 is required for the assembly of DRC8, and DRC8/11 form a subcomplex in the proximal lobe of the linker domain that is required to form stable contacts to the neighboring B-tubule. Gold labeling of tagged subunits determines the precise locations of the previously ambiguous N terminus of DRC4 and C terminus of DRC5. DRC4 is now shown to contribute to the core scaffold of the N-DRC. Our results reveal the overall architecture of N-DRC, with the 3 subunits DRC1/2/4 forming a core complex that serves as the scaffold for the assembly of the "functional subunits," namely DRC3/5-8/11. These findings shed light on N-DRC assembly and its role in regulating flagellar beating.

KW - Axoneme

KW - Cilia

KW - Cryo-electron tomography

KW - Flagella

KW - N-DRC

UR - http://www.scopus.com/inward/record.url?scp=85074926910&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85074926910&partnerID=8YFLogxK

U2 - 10.1073/pnas.1910960116

DO - 10.1073/pnas.1910960116

M3 - Article

C2 - 31659045

AN - SCOPUS:85074926910

VL - 116

SP - 23152

EP - 23162

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 46

ER -