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Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins
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Methods to analyze novel liaisons between endosomes and centrosomes
Hui-Fang Hung1, Heidi Hehnly2, Stephen Doxsey1
1Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA.
Methods in Cell Biology
|September 12, 2015
Summary
Recycling endosomes (REs) dynamically interact with the centrosome, influencing endosome recycling and centrosome maturation. This study reveals novel bidirectional roles between REs and the centrosome throughout the cell cycle.
Area of Science:
- Cell Biology
- Molecular Biology
- Cellular Dynamics
Background:
- Recycling endosomes (REs) are known to reside in the pericentrosomal region.
- The precise interactions between REs and the centrosome are not fully understood.
Purpose of the Study:
- To investigate the dynamic relationship between recycling endosomes and the centrosome.
- To uncover novel functions of the centrosome in REs and vice versa.
Main Methods:
- Centrosome isolation
- Live cell imaging
- Electron microscopy
Main Results:
- REs dynamically localize to the centrosome across the cell cycle.
- REs interact with the older centriole to regulate endosome recycling, involving the small GTPase Rab11.
- REs transport centrosome proteins to spindle poles during centrosome maturation.
Conclusions:
- The centrosome plays active roles in regulating RE function.
- REs contribute to centrosome function, specifically in centrosome maturation.
- This research highlights a novel, interdependent relationship between REs and the centrosome.
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