Actin filaments as dynamic reservoirs for Drp1 recruitment

Anna L Hatch; Wei-Ke Ji; Ronald A Merrill; Stefan Strack; Henry N Higgs

doi:10.1091/mbc.E16-03-0193

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Actin filaments as dynamic reservoirs for Drp1 recruitment

Journal article

Open access

Peer reviewed

Actin filaments as dynamic reservoirs for Drp1 recruitment

Anna L Hatch, Wei-Ke Ji, Ronald A Merrill, Stefan Strack and Henry N Higgs

Molecular biology of the cell, Vol.27(20), pp.3109-3121

10/15/2016

DOI: 10.1091/mbc.E16-03-0193

PMCID: PMC5063618

PMID: 27559132

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Abstract

Drp1 is a dynamin-family GTPase recruited to mitochondria and peroxisomes, where it oligomerizes and drives membrane fission. Regulation of mitochondrial Drp1 recruitment is not fully understood. We previously showed that Drp1 binds actin filaments directly, and actin polymerization is necessary for mitochondrial Drp1 oligomerization in mammals. Here we show the Drp1/actin interaction displays unusual properties that are influenced by several factors. At saturation, only a fraction Drp1 binds actin filaments, and the off-rate of actin-bound Drp1 is significantly increased by unbound Drp1. GDP and GTP accelerate and decelerate Drp1/actin binding dynamics, respectively. Actin has a biphasic effect on Drp1 GTP hydrolysis, increasing at low actin:Drp1 ratio but returning to baseline at high ratio. Drp1 also bundles filaments. Bundles have reduced dynamics but follow the same trends as single filaments. Drp1 preferentially incorporates into bundles at higher ionic strength. We measure Drp1 concentration to be ∼0.5 μM in U2OS cell cytosol, suggesting the actin-binding affinity measured here (K = 0.6 μM) is in the physiologically relevant range. The ability of Drp1 to bind actin filaments in a highly dynamic manner provides potential for actin filaments to serve as reservoirs of oligomerization-competent Drp1 that can be accessed for mitochondrial fission.

Actin Cytoskeleton - metabolism

GTP Phosphohydrolases - physiology

Microtubule-Associated Proteins - metabolism

Microtubule-Associated Proteins - pharmacokinetics

Humans

Protein Multimerization

Actins - metabolism

Dynamins

Mitochondrial Proteins - physiology

Mitochondria - metabolism

Peroxisomes - metabolism

GTP Phosphohydrolases - pharmacokinetics

Microtubule-Associated Proteins - physiology

Mitochondrial Dynamics - physiology

Hydrolysis

Animals

GTP Phosphohydrolases - metabolism

Mitochondrial Proteins - metabolism

Protein Binding

Cytosol - metabolism

Cell Culture Techniques

Mitochondrial Proteins - pharmacokinetics

Details

Title: Subtitle: Actin filaments as dynamic reservoirs for Drp1 recruitment
Creators: Anna L Hatch - Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth College, Hanover, NH 03755
Wei-Ke Ji - Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth College, Hanover, NH 03755
Ronald A Merrill - Department of Pharmacology, Carver School of Medicine, University of Iowa, Iowa City, IA 52242
Stefan Strack - Department of Pharmacology, Carver School of Medicine, University of Iowa, Iowa City, IA 52242
Henry N Higgs - Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth College, Hanover, NH 03755 henry.higgs@dartmouth.edu
Resource Type: Journal article
Publication Details: Molecular biology of the cell, Vol.27(20), pp.3109-3121
DOI: 10.1091/mbc.E16-03-0193
PMID: 27559132
PMCID: PMC5063618
NLM abbreviation: Mol Biol Cell
ISSN: 1059-1524
eISSN: 1939-4586
Publisher: American Society for Cell Biology; United States
Grant note: R56 NS056244 / NINDS NIH HHS R21 NS087908 / NINDS NIH HHS P20 GM113132 / NIGMS NIH HHS P30 CA023108 / NCI NIH HHS R01 GM106000 / NIGMS NIH HHS R01 GM069818 / NIGMS NIH HHS R01 NS056244 / NINDS NIH HHS
Language: English
Date published: 10/15/2016
Academic Unit: Pathology; Iowa Neuroscience Institute; Neuroscience and Pharmacology
Record Identifier: 9984040343802771

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