caa a22 4500 606190813 CHVBK 20210128100857.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s10577-015-9472-x doi (NATIONALLICENCE)springer-10.1007/s10577-015-9472-x RZZ and Mad1 dynamics in Drosophila mitosis [Elektronische Daten] [Lénaïg Défachelles, Natacha Raich, Régine Terracol, Xavier Baudin, Byron Williams, Michael Goldberg, Roger Karess] The presence or absence of Mad1 at kinetochores is a major determinant of spindle assembly checkpoint (SAC) activity, the surveillance mechanism that delays anaphase onset if one or more kinetochores remain unattached to spindle fibers. Among the factors regulating the levels of Mad1 at kinetochores is the Rod, Zw10, and Zwilch (RZZ) complex, which is required for Mad1 recruitment through a mechanism that remains unknown. The relative dynamics and interactions of Mad1 and RZZ at kinetochores have not been extensively investigated, although Mad1 has been reported to be stably recruited to unattached kinetochores. In this study, we directly compare Mad1-green fluorescent protein (GFP) turnover dynamics on unattached Drosophila kinetochores with that of RZZ, tagged either with GFP-Rod or GFP-Zw10. We find that nearly 40% of kinetochore-bound Mad1 has a significant dynamic component, turning over with a half-life of 12s. RZZ in contrast is essentially stable on unattached kinetochores. In addition, we report that a fraction of RZZ and Mad1 can co-immunoprecipitate, indicating that the genetically determined recruitment hierarchy (in which Mad1 depends on RZZ) may reflect a physical association of the two complexes. Springer Science+Business Media Dordrecht, 2015 Mad1 nationallicence Mad2 nationallicence RZZ nationallicence kinetochore nationallicence Zw10 nationallicence Rough Deal nationallicence SAC : Spindle assembly checkpoint nationallicence RZZ : Rough deal-Zw10-Zwilch nationallicence APC/C : Anaphase promoting complex/cyclosome nationallicence GFP : Green fluorescent protein nationallicence RFP : Red fluorescent protein nationallicence K-MT : Kinetochore microtubule nationallicence NEB : Nuclear envelope breakdown nationallicence IP : Immunoprecipitation nationallicence FRAP : Fluorescence recovery after photobleaching nationallicence Défachelles Lénaïg Sorbonne Paris Cité, Université Paris Diderot, Paris, France aut Raich Natacha Sorbonne Paris Cité, Université Paris Diderot, Paris, France aut Terracol Régine Sorbonne Paris Cité, Université Paris Diderot, Paris, France aut Baudin Xavier Sorbonne Paris Cité, Université Paris Diderot, Paris, France aut Williams Byron Department of Molecular Biology and Genetics, Cornell University, Biotechnology Building, 14853, Ithaca, NY, USA aut Goldberg Michael Department of Molecular Biology and Genetics, Cornell University, Biotechnology Building, 14853, Ithaca, NY, USA aut Karess Roger Sorbonne Paris Cité, Université Paris Diderot, Paris, France aut Chromosome Research Springer Netherlands 23/2(2015-06-01), 333-342 0967-3849 23:2<333 2015 23 10577 https://doi.org/10.1007/s10577-015-9472-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10577-015-9472-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Défachelles Lénaïg Sorbonne Paris Cité, Université Paris Diderot, Paris, France aut NATIONALLICENCE 700 1- Raich Natacha Sorbonne Paris Cité, Université Paris Diderot, Paris, France aut NATIONALLICENCE 700 1- Terracol Régine Sorbonne Paris Cité, Université Paris Diderot, Paris, France aut NATIONALLICENCE 700 1- Baudin Xavier Sorbonne Paris Cité, Université Paris Diderot, Paris, France aut NATIONALLICENCE 700 1- Williams Byron Department of Molecular Biology and Genetics, Cornell University, Biotechnology Building, 14853, Ithaca, NY, USA aut NATIONALLICENCE 700 1- Goldberg Michael Department of Molecular Biology and Genetics, Cornell University, Biotechnology Building, 14853, Ithaca, NY, USA aut NATIONALLICENCE 700 1- Karess Roger Sorbonne Paris Cité, Université Paris Diderot, Paris, France aut NATIONALLICENCE 773 0- Chromosome Research Springer Netherlands 23/2(2015-06-01), 333-342 0967-3849 23:2<333 2015 23 10577