title: A microtubule interactome: complexes with roles in cell cycle and mitosis.
abstract: The microtubule (MT) cytoskeleton is required for many aspects of cell function, including the transport of intracellular materials, the maintenance of cell polarity, and the regulation of mitosis. These functions are coordinated by MT-associated proteins (MAPs), which work in concert with each other, binding MTs and altering their properties. We have used a MT cosedimentation assay, combined with 1D and 2D PAGE and mass spectrometry, to identify over 250 MAPs from early Drosophila embryos. We have taken two complementary approaches to analyse the cellular function of novel MAPs isolated using this approach. First, we have carried out an RNA interference (RNAi) screen, identifying 21 previously uncharacterised genes involved in MT organisation. Second, we have undertaken a bioinformatics analysis based on binary protein interaction data to produce putative interaction networks of MAPs. By combining both approaches, we have identified and validated MAP complexes with potentially important roles in cell cycle regulation and mitosis. This study therefore demonstrates that biologically relevant data can be harvested using such a multidisciplinary approach, and identifies new MAPs, many of which appear to be important in cell division.
authors.affiliation: Department of Zoology, University of Oxford, Oxford, United Kingdom.
authors.forename: Julian R
authors.initials: JR
authors.lastname: Hughes
authors.forename: Ana M
authors.initials: AM
authors.lastname: Meireles
authors.forename: Katherine H
authors.initials: KH
authors.lastname: Fisher
authors.forename: Angel
authors.initials: A
authors.lastname: Garcia
authors.forename: Philip R
authors.initials: PR
authors.lastname: Antrobus
authors.forename: Alan
authors.initials: A
authors.lastname: Wainman
authors.forename: Nicole
authors.initials: N
authors.lastname: Zitzmann
authors.forename: Charlotte
authors.initials: C
authors.lastname: Deane
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Ohkura
authors.forename: James G
authors.initials: JG
authors.lastname: Wakefield
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D018385:Centrosome / Q000378:metabolism; D029721:Drosophila Proteins / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism*; D008938:Mitosis / Q000502:physiology*; D034622:RNA Interference; D044843:SKP Cullin F-Box Protein Ligases / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D008869:Microtubule-Associated Proteins; C434749:SkpA protein, Drosophila; D044843:SKP Cullin F-Box Protein Ligases