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Structure paper

TitleAxonemal dynein contributions to flagellar beat types and waveforms.
Journal, issue, pagesbioRxiv, Year 2025
Publish dateNov 7, 2025
AuthorsSophia Fochler / Matthew H Doran / Tom Beneke / James Smith / Cecile Fort / Benjamin J Walker / Alan Brown / Eva Gluenz / Richard J Wheeler /
PubMed AbstractEukaryotic flagella, or motile cilia, are iconic molecular machines whose beating drives cell propulsion and fluid transport across diverse organisms. Beat type and waveform are tailored to function, ...Eukaryotic flagella, or motile cilia, are iconic molecular machines whose beating drives cell propulsion and fluid transport across diverse organisms. Beat type and waveform are tailored to function, differing between species and cell types, and individual flagella can switch between beat types. Aberrant beating causes ciliopathies and infertility in humans and prevents unicellular parasite transmission. Eight distinct dynein motor protein complexes bind to axonemal doublet microtubules (DMTs) within flagella and drive beating, yet despite extensive structural analysis, how this machinery achieves different beat types is unknown. Here, using the flagellate unicellular parasite , we show a division of labour where specific dyneins drive specific beat types. Using cryo-EM, we determined the structure of the 96-nm repeat unit of the DMT and identified its dynein composition. We used CRISPR-Cas9 to systematically delete all 96-nm repeat proteins, comprehensively mapping necessity for swimming, and determined the contribution of each dynein to incidence and waveform of the preferred beat types. Outer dynein arms (ODAs) were required for symmetric tip-to-base beats, specific single-headed inner dynein arms (IDAs) were important for asymmetric base-to-tip beats (IDA), and double-headed IDA important for both. This systematic analysis indicates that the prevailing dogma that ODAs drive and IDAs shape the beat is either incomplete or not universal, and establishes new hypotheses for how different species, cell types and individual flagella achieve their necessary beat types.
External linksbioRxiv / PubMed:41279373 / PubMed Central
MethodsEM (single particle)
Resolution3.1 - 17.0 Å
Structure data

EMDB-71903: 96 nm half 1 consensus map
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-72431: Leishmania 96 nm half 1 composite map
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-72432: 96 nm half 2 composite map
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-72433: Leishmania 96 nm half 2 consensus map
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-72434: Structure of the Leishmania inner dyenin arm A base bound to the DMT
Method: EM (single particle) / Resolution: 3.6 Å

EMDB-72435: Structure of the Leishmania inner dyenin arm A motor domain
Method: EM (single particle) / Resolution: 9.0 Å

EMDB-72436: Structure of the Leishmania inner dyenin arm B base bound to the DMT
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72437: Structure of the Leishmania inner dyenin arm B motor domain
Method: EM (single particle) / Resolution: 9.0 Å

EMDB-72438: Structure of the Leishmania inner dyenin arm C base bound to the DMT
Method: EM (single particle) / Resolution: 4.0 Å

EMDB-72439: Structure of the Leishmania inner dyenin arm C motor domain
Method: EM (single particle) / Resolution: 17.0 Å

EMDB-72440: Structure of the Leishmania inner dyenin arm D and inner dynein arm G bases bound to the DMT
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-72441: Structure of the Leishmania inner dyenin arm D motor domain
Method: EM (single particle) / Resolution: 13.0 Å

EMDB-72442: Structure of the Leishmania inner dyenin arm E base bound to the DMT
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-72443: Structure of the Leishmania inner dyenin arm E motor domain
Method: EM (single particle) / Resolution: 15.0 Å

EMDB-72444: Structure of the Leishmania inner dyenin arm F base bound to the DMT
Method: EM (single particle) / Resolution: 8.1 Å

EMDB-72445: Structure of the Leishmania inner dyenin arm F motor domains
Method: EM (single particle) / Resolution: 7.0 Å

EMDB-72446: NDRC baseplate protofilament refinement
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-72447: Structure of the distal region of the Leishmania NDRC
Method: EM (single particle) / Resolution: 4.5 Å

EMDB-72448: Structure of the core of the Leishmania Outer Dynein Arm
Method: EM (single particle) / Resolution: 4.3 Å

EMDB-72449: Structure of the Leishmania Outer Dynein Arm motor domains
Method: EM (single particle) / Resolution: 4.8 Å

EMDB-72450: Structure of the Leishmania radial spoke 1 base bound to the DMT
Method: EM (single particle) / Resolution: 5.2 Å

EMDB-72451: Structure of the Leishmania radial spoke 2 base bound to the DMT
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-72452: Structure of the Leishmania radial spoke 3 base portion 1 bound to the DMT
Method: EM (single particle) / Resolution: 6.6 Å

EMDB-72453: Structure of the Leishmania radial spoke 3 portion 2 bound to the DMT
Method: EM (single particle) / Resolution: 6.7 Å

EMDB-72454: Structure of the Leishmania tether-tetherhead complex
Method: EM (single particle) / Resolution: 7.6 Å

EMDB-72563: Leishmania 96 nm half 1 protofilament refinement position 1_1
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-72564: 96 nm half 1 protofilament refinement position 1_2
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-72565: Leishmania 96 nm half 1 protofilament refinement position 1_3
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-72566: Leishmania 96 nm half 1 protofilament refinement position 2_1
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72567: Leishmania 96 nm half 1 protofilament refinement position 2_2
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72568: Leishmania 96 nm half 1 protofilament refinement position 2_3
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72569: Leishmania 96 nm half 1 protofilament refinement position 3_1
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72570: Leishmania 96 nm half 1 protofilament refinement position 3_2
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72571: Leishmania 96 nm half 1 protofilament refinement position 3_3
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72572: Leishmania 96 nm half 1 protofilament refinement position 4_1
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72573: Leishmania 96 nm half 1 protofilament refinement position 4_2
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72574: Leishmania 96 nm half 1 protofilament refinement position 4_3
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72575: Leishmania 96 nm half 1 protofilament refinement position 5_1
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72576: Leishmania 96 nm half 1 protofilament refinement position 5_2
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72577: Leishmania 96 nm half 1 protofilament refinement position 5_3
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72578: Leishmania 96 nm half 1 protofilament refinement position 6_1
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-72579: Leishmania 96 nm half 1 protofilament refinement position 6_2
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-72580: Leishmania 96 nm half 1 protofilament refinement position 6_3
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-72581: Leishmania 96 nm half 1 protofilament refinement position 7_1
Method: EM (single particle) / Resolution: 3.6 Å

EMDB-72582: Leishmania 96 nm half 1 protofilament refinement position 7_2
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72583: Leishmania 96 nm half 1 protofilament refinement position 7_3
Method: EM (single particle) / Resolution: 3.6 Å

EMDB-72584: Leishmania 96 nm half 1 protofilament refinement position 8_1
Method: EM (single particle) / Resolution: 3.6 Å

EMDB-72585: Leishmania 96 nm half 1 protofilament refinement position 8_2
Method: EM (single particle) / Resolution: 3.6 Å

EMDB-72586: Leishmania 96 nm half 1 protofilament refinement position 8_3
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72587: Leishmania 96 nm half 1 protofilament refinement position 9_1
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72588: Leishmania 96 nm half 1 protofilament refinement position 9_2
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72589: Leishmania 96 nm half 1 protofilament refinement position 9_3
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72590: Leishmania 96 nm half 1 protofilament refinement position 10_1
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72591: Leishmania 96 nm half 1 protofilament refinement position 10_2
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72592: Leishmania 96 nm half 1 protofilament refinement position 10_3
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72593: Leishmania 96 nm half 2 protofilament refinement position 1_1
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-72594: Leishmania 96 nm half 2 protofilament refinement position 1_2
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-72595: Leishmania 96 nm half 2 protofilament refinement position 1_3
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-72596: Leishmania 96 nm half 2 protofilament refinement position 2_1
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72597: Leishmania 96 nm half 2 protofilament refinement position 2_2
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-72598: Leishmania 96 nm half 2 protofilament refinement position 2_3
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-72599: Leishmania 96 nm half 2 protofilament refinement position 3_1
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72600: Leishmania 96 nm half 2 protofilament refinement position 3_2
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72601: Leishmania 96 nm half 2 protofilament refinement position 3_3
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72602: Leishmania 96 nm half 2 protofilament refinement position 4_1
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72603: Leishmania 96 nm half 2 protofilament refinement position 4_2
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72604: Leishmania 96 nm half 2 protofilament refinement position 4_3
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72605: Leishmania 96 nm half 2 protofilament refinement position 5_1
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72606: Leishmania 96 nm half 2 protofilament refinement position 5_2
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72607: Leishmania 96 nm half 2 protofilament refinement position 5_3
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72608: Leishmania 96 nm half 2 protofilament refinement position 6_1
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-72609: Leishmania 96 nm half 2 protofilament refinement position 6_2
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-72610: Leishmania 96 nm half 2 protofilament refinement position 6_3
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-72611: Leishmania 96 nm half 2 protofilament refinement position 7_1
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72612: Leishmania 96 nm half 2 protofilament refinement position 7_2
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72613: Leishmania 96 nm half 2 protofilament refinement position 7_3
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72614: Leishmania 96 nm half 2 protofilament refinement position 8_1
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72615: Leishmania 96 nm half 2 protofilament refinement position 8_2
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72616: Leishmania 96 nm half 2 protofilament refinement position 8_3
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-72617: Leishmania 96 nm half 2 protofilament refinement position 9_1
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72618: Leishmania 96 nm half 2 protofilament refinement position 9_2
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72619: Leishmania 96 nm half 2 protofilament refinement position 9_3
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-72620: Leishmania 96 nm half 2 protofilament refinement position 10_1
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-72621: Leishmania 96 nm half 2 protofilament refinement position 10_2
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-72622: Leishmania 96 nm half 2 protofilament refinement position 10_3
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-72629, PDB-9y6s:
96-nm repeat of the Leishmania tarentolae doublet microtubule
Method: EM (single particle) / Resolution: 3.7 Å

Chemicals

ChemComp-GDP:
GUANOSINE-5'-DIPHOSPHATE / GDP, energy-carrying molecule*YM

ChemComp-MG:
Unknown entry

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM

ChemComp-ZN:
Unknown entry

ChemComp-CA:
Unknown entry

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM

Source
  • leishmania tarentolae (eukaryote)
  • leishmania mexicana (eukaryote)
KeywordsSTRUCTURAL PROTEIN / flagella / parasite / axoneme / doublet microtubule

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