title: A maternal effect rough deal mutation suggests that multiple pathways regulate Drosophila RZZ kinetochore recruitment.
abstract: Proper kinetochore recruitment and regulation of dynein and the Mad1-Mad2 complex requires the Rod-Zw10-Zwilch (RZZ) complex. Here, we describe rod(Z3), a maternal-effect Drosophila mutation changing a single residue in the Rough Deal (Rod) subunit of RZZ. Although the RZZ complex containing this altered subunit (denoted R(Z3)ZZ) is present in early syncytial stage embryos laid by homozygous rod(Z3) mothers, it is not recruited to kinetochores. Consequently, the embryos have no spindle assembly checkpoint (SAC), and syncytial mitoses are profoundly perturbed. The polar body (residual meiotic products) cannot remain in its SAC-dependent metaphase-like state, and decondenses into chromatin. In neuroblasts of homozygous rod(Z3) larvae, R(Z3)ZZ recruitment is only partially reduced, the SAC is functional and mitosis is relatively normal. R(Z3)ZZ nevertheless behaves abnormally: it does not further accumulate on kinetochores when microtubules are depolymerized; it reduces the rate of Mad1 recruitment; and it dominantly interferes with the dynein-mediated streaming of RZZ from attached kinetochores. These results suggest that the mutated residue of rod(Z3) is required for normal RZZ kinetochore recruitment and function and, moreover, that the RZZ recruitment pathway might differ in syncytial stage embryos and post-embryonic somatic cells.
authors.affiliation: Equipe Labellisée Ligue Contre le Cancer, CNRS, Institut Jacques Monod, UMR7592, Université Paris Diderot, Sorbonne Paris Cité, Paris Cedex 13 75205, France.
authors.forename: Lénaïg
authors.initials: L
authors.lastname: Défachelles
authors.affiliation: Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232-8240, USA.
authors.forename: Sarah G
authors.initials: SG
authors.lastname: Hainline
authors.affiliation: Equipe Labellisée Ligue Contre le Cancer, CNRS, Institut Jacques Monod, UMR7592, Université Paris Diderot, Sorbonne Paris Cité, Paris Cedex 13 75205, France.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Menant
authors.affiliation: Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232-8240, USA.
authors.forename: Laura A
authors.initials: LA
authors.lastname: Lee
authors.affiliation: Equipe Labellisée Ligue Contre le Cancer, CNRS, Institut Jacques Monod, UMR7592, Université Paris Diderot, Sorbonne Paris Cité, Paris Cedex 13 75205, France karess.roger@ijm.univ-paris-diderot.fr.
authors.forename: Roger E
authors.initials: RE
authors.lastname: Karess
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000235:genetics* / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism; D005260:Female; D007091:Image Processing, Computer-Assisted; D018386:Kinetochores / Q000502:physiology*; D008869:Microtubule-Associated Proteins / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D008941:Spindle Apparatus / Q000502:physiology
chemical_list: D018797:Cell Cycle Proteins; D029721:Drosophila Proteins; D008869:Microtubule-Associated Proteins; C064993:ROD protein, Drosophila; C474841:Zwilch protein, Drosophila; C076055:Zw10 protein, Drosophila
keywords: Kinetochore; Mad1; Polar body; RZZ; Spindle assembly checkpoint; Syncytial embryo