title: Cryo-electron tomography reveals conserved features of doublet microtubules in flagella.
abstract: The axoneme forms the essential and conserved core of cilia and flagella. We have used cryo-electron tomography of Chlamydomonas and sea urchin flagella to answer long-standing questions and to provide information about the structure of axonemal doublet microtubules (DMTs). Solving an ongoing controversy, we show that B-tubules of DMTs contain exactly 10 protofilaments (PFs) and that the inner junction (IJ) and outer junction between the A- and B-tubules are fundamentally different. The outer junction, crucial for the initiation of doublet formation, appears to be formed by close interactions between the tubulin subunits of three PFs with unusual tubulin interfaces; other investigators have reported that this junction is weakened by mutations affecting posttranslational modifications of tubulin. The IJ consists of an axially periodic ladder-like structure connecting tubulin PFs of the A- and B-tubules. The recently discovered microtubule inner proteins (MIPs) on the inside of the A- and B-tubules are more complex than previously thought. They are composed of alternating small and large subunits with periodicities of 16 and/or 48 nm. MIP3 forms arches connecting B-tubule PFs, contrary to an earlier report that MIP3 forms the IJ. Finally, the "beak" structures within the B-tubules of Chlamydomonas DMT1, DMT5, and DMT6 are clearly composed of a longitudinal band of proteins repeating with a periodicity of 16 nm. These findings, discussed in relation to genetic and biochemical data, provide a critical foundation for future work on the molecular assembly and stability of the axoneme, as well as its function in motility and sensory transduction.
authors.affiliation: Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA 02454, USA. nicastro@brandeis.edu
authors.forename: Daniela
authors.initials: D
authors.lastname: Nicastro
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Fu
authors.forename: Thomas
authors.initials: T
authors.lastname: Heuser
authors.forename: Alan
authors.initials: A
authors.lastname: Tso
authors.forename: Mary E
authors.initials: ME
authors.lastname: Porter
authors.forename: Richard W
authors.initials: RW
authors.lastname: Linck
mesh_terms: D000818:Animals; D054468:Axoneme / Q000737:chemistry / Q000648:ultrastructure*; D002696:Chlamydomonas / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure; D020285:Cryoelectron Microscopy; D055032:Electron Microscope Tomography; D005407:Flagella / Q000737:chemistry / Q000648:ultrastructure*; D021621:Imaging, Three-Dimensional; D008297:Male; D008958:Models, Molecular; D010940:Plant Proteins / Q000737:chemistry; D021122:Protein Subunits; D013082:Sperm Tail / Q000737:chemistry / Q000648:ultrastructure; D047270:Strongylocentrotus purpuratus / Q000737:chemistry / Q000648:ultrastructure; D014404:Tubulin / Q000737:chemistry
chemical_list: D010940:Plant Proteins; D021122:Protein Subunits; D014404:Tubulin