An endosomal tether undergoes an entropic collapse to bring vesicles together

David H. Murray, Marcus Jahnel, Janelle Lauer, Mario J. Avellaneda, Nicolas Brouilly, Alice Cezanne, Hernán Morales-Navarrete, Enrico D. Perini, Charles Ferguson, Andrei N. Lupas, Yannis Kalaidzidis, Robert G. Parton, Stephan W. Grill (Lead / Corresponding author), Marino Zerial (Lead / Corresponding author)

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

An early step in intracellular transport is the selective recognition of a vesicle by its appropriate target membrane, a process regulated by Rab GTPases via the recruitment of tethering effectors. Membrane tethering confers higher selectivity and efficiency to membrane fusion than the pairing of SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) alone. Here we address the mechanism whereby a tethered vesicle comes closer towards its target membrane for fusion by reconstituting an endosomal asymmetric tethering machinery consisting of the dimeric coiled-coil protein EEA1 (refs 6, 7) recruited to phosphatidylinositol 3-phosphate membranes and binding vesicles harbouring Rab5. Surprisingly, structural analysis reveals that Rab5:GTP induces an allosteric conformational change in EEA1, from extended to flexible and collapsed. Through dynamic analysis by optical tweezers, we confirm that EEA1 captures a vesicle at a distance corresponding to its extended conformation, and directly measure its flexibility and the forces induced during the tethering reaction. Expression of engineered EEA1 variants defective in the conformational change induce prominent clusters of tethered vesicles in vivo. Our results suggest a new mechanism in which Rab5 induces a change in flexibility of EEA1, generating an entropic collapse force that pulls the captured vesicle towards the target membrane to initiate docking and fusion.

Original languageEnglish
Pages (from-to)107-111
Number of pages5
JournalNature
Volume537
Issue number7618
DOIs
Publication statusPublished - 24 Aug 2016

Fingerprint

Membrane Fusion
Membranes
rab GTP-Binding Proteins
Optical Tweezers
SNARE Proteins
Guanosine Triphosphate
Proteins
phosphatidylinositol 3-phosphate

Cite this

Murray, D. H., Jahnel, M., Lauer, J., Avellaneda, M. J., Brouilly, N., Cezanne, A., ... Zerial, M. (2016). An endosomal tether undergoes an entropic collapse to bring vesicles together. Nature, 537(7618), 107-111. https://doi.org/10.1038/nature19326
Murray, David H. ; Jahnel, Marcus ; Lauer, Janelle ; Avellaneda, Mario J. ; Brouilly, Nicolas ; Cezanne, Alice ; Morales-Navarrete, Hernán ; Perini, Enrico D. ; Ferguson, Charles ; Lupas, Andrei N. ; Kalaidzidis, Yannis ; Parton, Robert G. ; Grill, Stephan W. ; Zerial, Marino. / An endosomal tether undergoes an entropic collapse to bring vesicles together. In: Nature. 2016 ; Vol. 537, No. 7618. pp. 107-111.
@article{00a95a395a194966be7972040a5070e3,
title = "An endosomal tether undergoes an entropic collapse to bring vesicles together",
abstract = "An early step in intracellular transport is the selective recognition of a vesicle by its appropriate target membrane, a process regulated by Rab GTPases via the recruitment of tethering effectors. Membrane tethering confers higher selectivity and efficiency to membrane fusion than the pairing of SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) alone. Here we address the mechanism whereby a tethered vesicle comes closer towards its target membrane for fusion by reconstituting an endosomal asymmetric tethering machinery consisting of the dimeric coiled-coil protein EEA1 (refs 6, 7) recruited to phosphatidylinositol 3-phosphate membranes and binding vesicles harbouring Rab5. Surprisingly, structural analysis reveals that Rab5:GTP induces an allosteric conformational change in EEA1, from extended to flexible and collapsed. Through dynamic analysis by optical tweezers, we confirm that EEA1 captures a vesicle at a distance corresponding to its extended conformation, and directly measure its flexibility and the forces induced during the tethering reaction. Expression of engineered EEA1 variants defective in the conformational change induce prominent clusters of tethered vesicles in vivo. Our results suggest a new mechanism in which Rab5 induces a change in flexibility of EEA1, generating an entropic collapse force that pulls the captured vesicle towards the target membrane to initiate docking and fusion.",
author = "Murray, {David H.} and Marcus Jahnel and Janelle Lauer and Avellaneda, {Mario J.} and Nicolas Brouilly and Alice Cezanne and Hern{\'a}n Morales-Navarrete and Perini, {Enrico D.} and Charles Ferguson and Lupas, {Andrei N.} and Yannis Kalaidzidis and Parton, {Robert G.} and Grill, {Stephan W.} and Marino Zerial",
year = "2016",
month = "8",
day = "24",
doi = "10.1038/nature19326",
language = "English",
volume = "537",
pages = "107--111",
journal = "Nature",
issn = "0028-0836",
publisher = "Nature Publishing Group",
number = "7618",

}

Murray, DH, Jahnel, M, Lauer, J, Avellaneda, MJ, Brouilly, N, Cezanne, A, Morales-Navarrete, H, Perini, ED, Ferguson, C, Lupas, AN, Kalaidzidis, Y, Parton, RG, Grill, SW & Zerial, M 2016, 'An endosomal tether undergoes an entropic collapse to bring vesicles together', Nature, vol. 537, no. 7618, pp. 107-111. https://doi.org/10.1038/nature19326

An endosomal tether undergoes an entropic collapse to bring vesicles together. / Murray, David H.; Jahnel, Marcus; Lauer, Janelle; Avellaneda, Mario J.; Brouilly, Nicolas; Cezanne, Alice; Morales-Navarrete, Hernán; Perini, Enrico D.; Ferguson, Charles; Lupas, Andrei N.; Kalaidzidis, Yannis; Parton, Robert G.; Grill, Stephan W. (Lead / Corresponding author); Zerial, Marino (Lead / Corresponding author).

In: Nature, Vol. 537, No. 7618, 24.08.2016, p. 107-111.

Research output: Contribution to journalArticle

TY - JOUR

T1 - An endosomal tether undergoes an entropic collapse to bring vesicles together

AU - Murray, David H.

AU - Jahnel, Marcus

AU - Lauer, Janelle

AU - Avellaneda, Mario J.

AU - Brouilly, Nicolas

AU - Cezanne, Alice

AU - Morales-Navarrete, Hernán

AU - Perini, Enrico D.

AU - Ferguson, Charles

AU - Lupas, Andrei N.

AU - Kalaidzidis, Yannis

AU - Parton, Robert G.

AU - Grill, Stephan W.

AU - Zerial, Marino

PY - 2016/8/24

Y1 - 2016/8/24

N2 - An early step in intracellular transport is the selective recognition of a vesicle by its appropriate target membrane, a process regulated by Rab GTPases via the recruitment of tethering effectors. Membrane tethering confers higher selectivity and efficiency to membrane fusion than the pairing of SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) alone. Here we address the mechanism whereby a tethered vesicle comes closer towards its target membrane for fusion by reconstituting an endosomal asymmetric tethering machinery consisting of the dimeric coiled-coil protein EEA1 (refs 6, 7) recruited to phosphatidylinositol 3-phosphate membranes and binding vesicles harbouring Rab5. Surprisingly, structural analysis reveals that Rab5:GTP induces an allosteric conformational change in EEA1, from extended to flexible and collapsed. Through dynamic analysis by optical tweezers, we confirm that EEA1 captures a vesicle at a distance corresponding to its extended conformation, and directly measure its flexibility and the forces induced during the tethering reaction. Expression of engineered EEA1 variants defective in the conformational change induce prominent clusters of tethered vesicles in vivo. Our results suggest a new mechanism in which Rab5 induces a change in flexibility of EEA1, generating an entropic collapse force that pulls the captured vesicle towards the target membrane to initiate docking and fusion.

AB - An early step in intracellular transport is the selective recognition of a vesicle by its appropriate target membrane, a process regulated by Rab GTPases via the recruitment of tethering effectors. Membrane tethering confers higher selectivity and efficiency to membrane fusion than the pairing of SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) alone. Here we address the mechanism whereby a tethered vesicle comes closer towards its target membrane for fusion by reconstituting an endosomal asymmetric tethering machinery consisting of the dimeric coiled-coil protein EEA1 (refs 6, 7) recruited to phosphatidylinositol 3-phosphate membranes and binding vesicles harbouring Rab5. Surprisingly, structural analysis reveals that Rab5:GTP induces an allosteric conformational change in EEA1, from extended to flexible and collapsed. Through dynamic analysis by optical tweezers, we confirm that EEA1 captures a vesicle at a distance corresponding to its extended conformation, and directly measure its flexibility and the forces induced during the tethering reaction. Expression of engineered EEA1 variants defective in the conformational change induce prominent clusters of tethered vesicles in vivo. Our results suggest a new mechanism in which Rab5 induces a change in flexibility of EEA1, generating an entropic collapse force that pulls the captured vesicle towards the target membrane to initiate docking and fusion.

UR - http://www.scopus.com/inward/record.url?scp=84984653623&partnerID=8YFLogxK

U2 - 10.1038/nature19326

DO - 10.1038/nature19326

M3 - Article

C2 - 27556945

AN - SCOPUS:84984653623

VL - 537

SP - 107

EP - 111

JO - Nature

JF - Nature

SN - 0028-0836

IS - 7618

ER -

Murray DH, Jahnel M, Lauer J, Avellaneda MJ, Brouilly N, Cezanne A et al. An endosomal tether undergoes an entropic collapse to bring vesicles together. Nature. 2016 Aug 24;537(7618):107-111. https://doi.org/10.1038/nature19326