Accéder au contenu
MilliporeSigma

The kinetochore-dependent and -independent formation of the CDC20-MAD2 complex and its functions in HeLa cells.

Scientific reports (2017-01-24)
Jianquan Li, Nanmao Dang, Daniel James Wood, Jun-Yong Huang
RÉSUMÉ

The mitotic checkpoint complex (MCC) is formed from two sub-complexes of CDC20-MAD2 and BUBR1-BUB3, and current models suggest that it is generated exclusively by the kinetochores after nuclear envelope breakdown (NEBD). However, neither sub-complex has been visualised in vivo, and when and where they are formed during the cell cycle and their response to different SAC conditions remains elusive. Using single cell analysis in HeLa cells, we show that the CDC20-MAD2 complex is cell cycle regulated with a "Bell" shaped profile and peaks at prometaphase. Its formation begins in early prophase before NEBD when the SAC has not been activated. The complex prevents the premature degradation of cyclin B1. Tpr, a component of the NPCs (nuclear pore complexes), facilitates the formation of this prophase form of the CDC20-MAD2 complex but is inactive later in mitosis. Thus, we demonstrate that the CDC20-MAD2 complex could also be formed independently of the SAC. Moreover, in prolonged arrest caused by nocodazole treatment, the overall levels of the CDC20-MAD2 complex are gradually, but significantly, reduced and this is associated with lower levels of cyclin B1, which brings a new insight into the mechanism of mitotic "slippage" of the arrested cells.

MATÉRIAUX
Référence du produit
Marque
Description du produit

Sigma-Aldrich
Réactifs de détection in situ Duolink® Red
Sigma-Aldrich
Sonde PLA® PLUS anti-lapin in situ Duolink®, Affinity purified Donkey anti-Rabbit IgG (H+L)
Sigma-Aldrich
Sonde PLA® in situ DuolinkDuolink® anti-souris MINUS, Affinity purified Donkey anti-Mouse IgG (H+L)
Sigma-Aldrich
Starter Kit Duolink® In Situ rouge - souris/lapin
Sigma-Aldrich
Réactifs de détection in situ Duolink® Orange
Sigma-Aldrich
Starter Kit Duolink® In Situ orange - souris/lapin
Sigma-Aldrich
Kit de réactifs de détection in situ dans le rouge lointain Duolink®
Sigma-Aldrich
Sonde PLA® in situ DuolinkDuolink® anti-lapin MINUS
Sigma-Aldrich
Réactifs de détection in situ Duolink® vert
Sigma-Aldrich
Duolink® In Situ Wash Buffers, Fluorescence
Sigma-Aldrich
Sonde PLA® in situ Duolink® anti-souris positive
Sigma-Aldrich
Duolink® In Situ Mounting Medium with DAPI
Sigma-Aldrich
CelLytic MT Cell Lysis Reagent, For mammalian tissues
Sigma-Aldrich
Kit de réactifs de détection in situ en fond clair Duolink®
Sigma-Aldrich
Z-Leu-Leu-Leu-al, ≥90% (HPLC)
Sigma-Aldrich
Duolink® In Situ Probemaker PLUS
Sigma-Aldrich
Duolink® In Situ Probemaker MINUS
Sigma-Aldrich
Duolink® In Situ PLA® Probe Anti-Goat MINUS, Affinity purified Donkey anti-Goat IgG (H+L)
Sigma-Aldrich
Duolink® PLA Control Kit - PPI, Control Kit to reliably detect Protein-Protein Interactions (PPI) with Duolink® PLA
Sigma-Aldrich
Duolink® In Situ PLA® Probe Anti-Goat PLUS
Sigma-Aldrich
Duolink® flowPLA Detection Kit - FarRed, Duolink® PLA kit for Flow Cytometry with FarRed Detection
Sigma-Aldrich
Duolink® flowPLA Detection Kit - Red, DUOLINK® : PLA kit for Flow Cytometry with Red Detection
Sigma-Aldrich
Duolink® In Situ Red Starter Kit Goat/Rabbit
Sigma-Aldrich
Duolink® flowPLA Detection Kit - Green, Duolink® PLA kit for Flow Cytometry with Green Detection
Sigma-Aldrich
Duolink® In Situ Wash Buffer, Brightfield
Sigma-Aldrich
Duolink® In Situ Microplate Nuclear Stain, Anti-Fade
Sigma-Aldrich
Duolink® flowPLA Detection Kit - Orange, Duolink® PLA kit for Flow Cytometry with Orange Detection
Sigma-Aldrich
Duolink® In Situ Microplate Heat Transfer Block
Sigma-Aldrich
Duolink® In Situ Red Starter Kit Mouse/Goat
Sigma-Aldrich
Duolink® In Situ Orange Starter Kit Goat/Rabbit