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Endosome Fusion and Tubule Formation in Cells Overexpressing GFP-Rab5 Fusion Proteins

Published online by Cambridge University Press:  02 July 2020

R.L. Roberts ,
M.A. Barbieri  and
P.D. Stahl
R.L. Roberts
Affiliation:
Department of Cell Biology and Physiology, Washington University, School of Medicine, St. Louis, MO., 63110.
M.A. Barbieri
Affiliation:
Department of Cell Biology and Physiology, Washington University, School of Medicine, St. Louis, MO., 63110.
P.D. Stahl
Affiliation:
Department of Cell Biology and Physiology, Washington University, School of Medicine, St. Louis, MO., 63110.
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Extract

Previous studies have demonstrated that GTPgS and rab5:Q79L, a constitutively active rab5 mutant, stimulate endosome fusion in in vitro assays [Cell Reg. 1:113-124 (1989)]. Additionally, electron microscopy has shown that under these conditions, endosomes form incompletely fused couplets [Cell Reg. 1:113-124 (1989); Arch. Biochem. Biophys. 326:64-72 (1996)]. CHO and BHK cells that overexpress rab5:Q79L develop ‘giant’ cytoplasmic vesicles that exhibit many characteristics of early endosomes including immunoreactivity for rab 5 and transferrin receptor (data not shown). The formation of these vesicles has been analyzed by time-lapse video microscopy which shows the enlarged endosomes arise primarily by fusion of smaller vesicles. These endosome fusion events occur by two distinct mechanisms that differ in temporal and spatial characteristics of the membrane merger process and we refer to these processes as “explosive” versus “bridge” fusion. “Explosive” fusion is a rapid process in which the initial opening that forms between contacting vesicles expands rapidly and permits an abrupt, complete coalescence of membranes (Fig. 1).

Type
Cell Biology Applications of Green Fluorescent Protein and Other Vital Labeling Probes
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 137 - 138
Copyright
Copyright © Microscopy Society of America 1997

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References

1.Mayorga, L.S.et al., Cell Reg. 1, 113–124.10.1091/mbc.1.1.113CrossRefGoogle Scholar
2.Barbieri, M.A.et al., Arch. Biochem. Biophys. 326: 64–72.10.1006/abbi.1996.0047CrossRefGoogle Scholar