Live-cell super-resolution microscopy reveals a primary role for diffusion in polyglutamine-driven aggresome assembly.

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cam.issuedOnline2018-11-06
dc.contributor.authorLu, Meng
dc.contributor.authorBanetta, Luca
dc.contributor.authorYoung, Laurence J
dc.contributor.authorSmith, Edward J
dc.contributor.authorBates, Gillian P
dc.contributor.authorZaccone, Alessio
dc.contributor.authorKaminski Schierle, Gabriele S
dc.contributor.authorTunnacliffe, Alan
dc.contributor.authorKaminski, Clemens F
dc.contributor.orcidLu, Meng [0000-0001-9311-2666]
dc.contributor.orcidBanetta, Luca [0000-0003-2055-9584]
dc.contributor.orcidKaminski Schierle, Gabriele [0000-0002-1843-2202]
dc.contributor.orcidKaminski, Clemens [0000-0002-5194-0962]
dc.date.accessioned2018-12-19T00:30:50Z
dc.date.available2018-12-19T00:30:50Z
dc.date.issued2019-01-04
dc.description.abstractThe mechanisms leading to self-assembly of misfolded proteins into amyloid aggregates have been studied extensively in the test tube under well-controlled conditions. However, to what extent these processes are representative of those in the cellular environment remains unclear. Using super-resolution imaging of live cells, we show here that an amyloidogenic polyglutamine-containing protein first forms small, amorphous aggregate clusters in the cytosol, chiefly by diffusion. Dynamic interactions among these clusters limited their elongation and led to structures with a branched morphology, differing from the predominantly linear fibrils observed in vitro Some of these clusters then assembled via active transport at the microtubule-organizing center and thereby initiated the formation of perinuclear aggresomes. Although it is widely believed that aggresome formation is entirely governed by active transport along microtubules, here we demonstrate, using a combined approach of advanced imaging and mathematical modeling, that diffusion is the principal mechanism driving aggresome expansion. We found that the increasing surface area of the expanding aggresome increases the rate of accretion caused by diffusion of cytosolic aggregates and that this pathway soon dominates aggresome assembly. Our findings lead to a different view of aggresome formation than that proposed previously. We also show that aggresomes mature over time, becoming more compacted as the structure grows. The presence of large perinuclear aggregates profoundly affects the behavior and health of the cell, and our super-resolution imaging results indicate that aggresome formation and development are governed by highly dynamic processes that could be important for the design of potential therapeutic strategies.
dc.format.mediumPrint-Electronic
dc.identifier.doi10.17863/CAM.34478
dc.identifier.eissn1083-351X
dc.identifier.issn0021-9258
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/287169
dc.languageeng
dc.publisherElsevier BV
dc.publisher.urlhttp://dx.doi.org/10.1074/jbc.ra118.003500
dc.subjectaggresome formation
dc.subjectamyloid protein
dc.subjectlive cell SIM
dc.subjectmolecular dynamics
dc.subjectmolecular imaging
dc.subjectmolecular modeling
dc.subjectpassive transport
dc.subjectprotein aggregation
dc.subjectprotein misfolding
dc.subjecttransport
dc.subjectAnimals
dc.subjectCell Nucleus
dc.subjectFemale
dc.subjectMale
dc.subjectMice
dc.subjectMicroscopy, Fluorescence
dc.subjectMicrotubule-Organizing Center
dc.subjectModels, Biological
dc.subjectPeptides
dc.titleLive-cell super-resolution microscopy reveals a primary role for diffusion in polyglutamine-driven aggresome assembly.
dc.typeArticle
dcterms.dateAccepted2018-10-29
prism.endingPage268
prism.issueIdentifier1
prism.publicationDate2019
prism.publicationNameJ Biol Chem
prism.startingPage257
prism.volume294
pubs.funder-project-idWellcome Trust (089703/Z/09/Z)
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/H023917/1)
pubs.funder-project-idEngineering and Physical Sciences Research Council (EP/H018301/1)
pubs.funder-project-idMedical Research Council (MR/K02292X/1)
pubs.funder-project-idMedical Research Council (G0902243)
pubs.funder-project-idMedical Research Council (MR/K015850/1)
pubs.funder-project-idEngineering and Physical Sciences Research Council (EP/L015889/1)
pubs.funder-project-idEuropean Research Council (233232)
pubs.funder-project-idWellcome Trust (203249/Z/16/Z)
rioxxterms.licenseref.startdate2019-01
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.typeJournal Article/Review
rioxxterms.versionAM
rioxxterms.versionofrecord10.1074/jbc.RA118.003500
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