Structural and geometrical constraints on the outer dynein arm in situ

Kurt Barkalow, Jock Avolio, Michael E.J. Holwill, Toshikazu Hamasaki, Peter Satir

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

This study considers the relationship between two structural forms of the 22S dynein arm of Tetrahymena thermophila: the bouquet and the compact arm. The compact arm differs from the bouquet and from other proposed forms (e.g., the “toadstool”) in that the globular domains are situated transversely across the interdoublet gap with one globular subunit, the head, proximal to the adjacent doublet microtubule. The other models place all three globular domains proximal to the neighboring doublet microtubule. When sliding of an isolated axoneme is induced, at least 57% of total attached arms on exposed doublets are in the compact form within dimensions of 24 × 24 × 12 nm, and only about 2% of the arms are bouquets. Toadstools are incompatible with the images seen. Bouquets are not found in regions of the doublet protected by a neighboring doublet. When axonemes with exposed doublets are treated with 0.5 M KCl for 30 min, the compct arms and the dynein heavy (H)‐chains disappear, while isolated bouquets and dynein H‐chains appear in the medium, suggesting that the compact arms give rise to the bouquets as they are solubilized. The bouquet is the predominant form of isolated 22S dynein molecules, which are found in two apparently enantiomorphic forms, within dimensions 45 × 39 × 13 nm; bouquets attached to doublets have dimensions similar to those of isolated bouquets. Computer modeling indicates that in an intact standard‐diameter axoneme, these dimensions are incompatible with the interdoublet volume available for an arm; the bouquet therefore represents an unfolded compact arm. A plausible sequence of changes can be modeled to illustrate the conversion of an attached compact arm to an attached and then free bouquet. The toadstool is probably an artifact that arises after unfolding. Consistent with the conformational difference, H‐chains of attached compact arms differ from those of isolated bouquets in their susceptibility to limited proteolysis. These results suggest that the compact arm, rather than the unfolded bouquet or the toadstool, is the functional form of the outer arm in the intact axoneme. © 1994 Wiley‐Liss, Inc.

Original languageEnglish (US)
Pages (from-to)299-312
Number of pages14
JournalCell motility and the cytoskeleton
Volume27
Issue number4
DOIs
StatePublished - 1994

Keywords

  • axoneme
  • cilia
  • computer modeling
  • dynein
  • microtubule motors

ASJC Scopus subject areas

  • Structural Biology
  • Cell Biology

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