Hemi-fused structure mediates and controls fusion and fission in live cells.
Membrane fusion and fission are vital for eukaryotic life. For three decades, it has been proposed that fusion is mediated by fusion between the proximal leaflets of two bilayers (hemi-fusion) to produce a hemi-fused structure, followed by fusion between the distal leaflets, whereas fission is via hemi-fission, which also produces a hemi-fused structure, followed by full fission. This hypothesis remained unsupported owing to the lack of observation of hemi-fusion or hemi-fission in live cells. A competing fusion hypothesis involving protein-lined pore formation has also been proposed. Here we report the observation of a hemi-fused Ω-shaped structure in live neuroendocrine chromaffin cells and pancreatic β-cells, visualized using confocal and super-resolution stimulated emission depletion microscopy. This structure is generated from fusion pore opening or closure (fission) at the plasma membrane. Unexpectedly, the transition to full fusion or fission is determined by competition between fusion and calcium/dynamin-dependent fission mechanisms, and is notably slow (seconds to tens of seconds) in a substantial fraction of the events. These results provide key missing evidence in support of the hemi-fusion and hemi-fission hypothesis in live cells, and reveal the hemi-fused intermediate as a key structure controlling fusion and fission, as fusion and fission mechanisms compete to determine the transition to fusion or fission.
National Institute of Neurological Disorders and Stroke, 35 Convent Drive, Room 2B-1012, Bethesda, Maryland 20892, USA.
Wei-Dong
WD
Zhao
National Institute of Neurological Disorders and Stroke, 35 Convent Drive, Room 2B-1012, Bethesda, Maryland 20892, USA.
Edaeni
E
Hamid
National Institute of Neurological Disorders and Stroke, 35 Convent Drive, Room 2B-1012, Bethesda, Maryland 20892, USA.
Wonchul
W
Shin
National Institute of Neurological Disorders and Stroke, 35 Convent Drive, Room 2B-1012, Bethesda, Maryland 20892, USA.
Peter J
PJ
Wen
National Institute on Deafness and other Communication Disorders, 35A Convent Drive, Room 3D-824, Bethesda, Maryland 20892, USA.
Evan S
ES
Krystofiak
National Institute of Neurological Disorders and Stroke, 35 Convent Drive, Room 2B-1012, Bethesda, Maryland 20892, USA.
Seth A
SA
Villarreal
National Institute of Neurological Disorders and Stroke, 35 Convent Drive, Room 2B-1012, Bethesda, Maryland 20892, USA.
Hsueh-Cheng
HC
Chiang
National Institute on Deafness and other Communication Disorders, 35A Convent Drive, Room 3D-824, Bethesda, Maryland 20892, USA.
Bechara
B
Kachar
National Institute of Neurological Disorders and Stroke, 35 Convent Drive, Room 2B-1012, Bethesda, Maryland 20892, USA.
Ling-Gang
LG
Wu
D000818:Animals; D001667:Binding, Competitive; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002470:Cell Survival; D002478:Cells, Cultured; D019439:Chromaffin Cells / Q000166:cytology; D034281:Dynamins / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000166:cytology; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism*; D008561:Membrane Fusion / Q000502:physiology*; D018613:Microscopy, Confocal; D008954:Models, Biological; D015203:Reproducibility of Results; D013997:Time Factors
D008051:Lipid Bilayers; D034281:Dynamins; D002118:Calcium