The Architecture of Outer Dynein Arms in Situ

Ishikawa, Takashi; Sakakibara, Hitoshi; Oiwa, Kazuhiro

doi:10.1016/j.jmb.2007.02.072

Cited by 113 publications

(109 citation statements)

References 59 publications

Supporting

Mentioning

100

Contrasting

Order By: Relevance

“…Cryo-TEM micrographs were recorded according to an established protocol. 35 Unilamellar and multilamellar vesicles with nonspherical morphology were indeed found (see Figure 2). Unfortunately, neither Lad-PC-Lad (1) nor Mad-PC-Mad (2) could be easily formulated into liposomes, as the suspensions were not stable, and a precipitate formed, a tendency also observed by DLS measurements in a smaller scale for Pad-PC-Pad vesicles during the course of several days.…”

Section: ■ Results and Discussionmentioning

confidence: 84%

Bilayer Properties of 1,3-Diamidophospholipids

et al. 2015

Self Cite

View full text Add to dashboard Cite

A series of 1,3-diamido phosphocholines was synthesized, and their potential to form stable bilayers was investigated. Large and giant unilamellar vesicles produced from these new lipids form a wide variety of faceted liposomes. Factors such as cooling rates and the careful choice of the liposome preparation method influence the formation of facets. Interdigitation was hypothesized as a main factor for the stabilization of facets and effectively monitored by small-angle X-ray scattering measurements.

show abstract

Section: ■ Results and Discussionmentioning

confidence: 84%

Bilayer Properties of 1,3-Diamidophospholipids

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…Human diseases due to ciliary motility defects [termed primary ciliary dyskinesia (PCD)] are caused most commonly by defects in OAD assembly (9)(10)(11). The assembly process and the in situ structure of the OAD complex in the axoneme have been well studied (3,12,13). However, the mechanism underlying the periodic binding of OAD to the doublet is poorly understood.…”

mentioning

confidence: 99%

“…In the flagella of Chlamydomonas mutants (e.g., outerdynein-arm deficient oda6) retaining the ODA-DC but not OAD, the ODA-DC is observed by electron microscopy as a small projection linearly arrayed every 24 nm along the outer doublet (3,(14)(15)(16)(17). It is composed of three subunits: DC1, ∼83 kDa, encoded by ODA3; DC2, ∼62 kDa, encoded by ODA1; and DC3, ∼21 kDa, encoded by ODA14 (18)(19)(20), which assemble in the cell cytoplasm and are transported into the flagella independently of OAD (16).…”

mentioning

confidence: 99%

Cooperative binding of the outer arm-docking complex underlies the regular arrangement of outer arm dynein in the axoneme

Owa

Furuta

Usukura

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Outer arm dynein (OAD) in cilia and flagella is bound to the outer doublet microtubules every 24 nm. Periodic binding of OADs at specific sites is important for efficient cilia/flagella beating; however, the molecular mechanism that specifies OAD arrangement remains elusive. Studies using the green alga Chlamydomonas reinhardtii have shown that the OAD-docking complex (ODA-DC), a heterotrimeric complex present at the OAD base, functions as the OAD docking site on the doublet. We find that the ODA-DC has an ellipsoidal shape ∼24 nm in length. In mutant axonemes that lack OAD but retain the ODA-DC, ODA-DC molecules are aligned in an end-to-end manner along the outer doublets. When flagella of a mutant lacking ODA-DCs are supplied with ODA-DCs upon gamete fusion, ODA-DC molecules first bind to the mutant axonemes in the proximal region, and the occupied region gradually extends toward the tip, followed by binding of OADs. This and other results indicate that a cooperative association of the ODA-DC underlies its function as the OAD-docking site and is the determinant of the 24-nm periodicity.C ilia and flagella of eukaryotic cells are organelles that generate fluid flow on the cell surface and/or sense chemical or mechanical stimuli from the external environment (1). Cilia/flagella beating is driven by outer arm dynein (OAD) and inner arm dyneins. The arrangement of dyneins on the axoneme has an overall periodicity of 96 nm, within which OAD binds every 24 nm; this 24-nm periodicity is completely conserved in essentially all eukaryotic organisms with "9 + 2" axonemes (2, 3) and even occurs in insect sperm flagella containing multiple rows of doublet microtubules arranged in a spiral configuration (4, 5). OAD is best characterized in Chlamydomonas. It is a very large protein complex of ∼2 MDa, comprising 3 heavy chains, 2 intermediate chains, and 11 distinct light chains. Most of the subunits are conserved from protists to mammals (6). OAD is the most abundant and most powerful axonemal dynein, generating about twothirds of the total propulsive force of ciliary beating (7,8). Human diseases due to ciliary motility defects [termed primary ciliary dyskinesia (PCD)] are caused most commonly by defects in OAD assembly (9-11). The assembly process and the in situ structure of the OAD complex in the axoneme have been well studied (3, 12, 13). However, the mechanism underlying the periodic binding of OAD to the doublet is poorly understood.The outer dynein arm-docking complex (ODA-DC) has been identified as a complex that mediates OAD binding to the doublet (14, 15). In the flagella of Chlamydomonas mutants (e.g., outerdynein-arm deficient oda6) retaining the ODA-DC but not OAD, the ODA-DC is observed by electron microscopy as a small projection linearly arrayed every 24 nm along the outer doublet (3,(14)(15)(16)(17). It is composed of three subunits: DC1, ∼83 kDa, encoded by ODA3; DC2, ∼62 kDa, encoded by ODA1; and DC3, ∼21 kDa, encoded by ODA14 (18-20), which assemble in the cell cytoplasm and are transported into the fla...

show abstract

“…Therefore, how dynein interacts with microtubules and how the observed conformational change in dynein contributes to the movement along microtubules remain unresolved questions. Although cryo-EM has produced 3D structures of axonemal dyneins (15)(16)(17), the head domains were not sufficiently well resolved to identify the sidedness or the polarity of the AAAϩ ring. Therefore, it was not possible to relate the conformational change observed in projection to the structure of dynein in the axoneme.…”

mentioning

confidence: 99%

Three-dimensional structure of cytoplasmic dynein bound to microtubules

Mizuno

Narita

Kon

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Cytoplasmic dynein is a large, microtubule-dependent molecular motor (1.2 MDa). Although the structure of dynein by itself has been characterized, its conformation in complex with microtubules is still unknown. Here, we used cryoelectron microscopy (cryo-EM) to visualize the interaction between dynein and microtubules. Most dynein molecules in the nucleotide-free state are bound to the microtubule in a defined conformation and orientation. A 3D image reconstruction revealed that dynein's head domain, formed by a ring-like arrangement of AAA؉ domains, is located Ϸ280 Å away from the center of the microtubule. The order of the AAA؉ domains in the ring was determined by using recombinant markers. Furthermore, a 3D helical image reconstruction of microtubules with a dynein's microtubule binding domain [dynein stalk (DS)] revealed that the stalk extends perpendicular to the microtubule. By combining the 3D maps of the dynein-microtubule and DSmicrotubule complexes, we present a model for how dynein in the nucleotide-free state binds to microtubules and discuss models for dynein's power stroke.cryoelectron microscopy ͉ Dictostelium

show abstract

The Architecture of Outer Dynein Arms in Situ

Cited by 113 publications

References 59 publications

Bilayer Properties of 1,3-Diamidophospholipids

Bilayer Properties of 1,3-Diamidophospholipids

Cooperative binding of the outer arm-docking complex underlies the regular arrangement of outer arm dynein in the axoneme

Three-dimensional structure of cytoplasmic dynein bound to microtubules

Contact Info

Product

Resources

About