Tunable microsecond dynamics of an allosteric switch regulate the activity of a AAA+ disaggregation machine

Hisham Mazal; Marija Iljina; Yoav Barak; Nadav Elad; Rina Rosenzweig; Pierre Goloubinoff; Inbal Riven; Gilad Haran

doi:10.1038/s41467-019-09474-6

Tunable microsecond dynamics of an allosteric switch regulate the activity of a AAA+ disaggregation machine

Hisham Mazal, Marija Iljina, Yoav Barak, Nadav Elad, Rina Rosenzweig, Pierre Goloubinoff, Inbal Riven, Gilad Haran^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

33 Citations (Scopus)

54 Downloads (Pure)

Abstract

Large protein machines are tightly regulated through allosteric communication channels. Here we demonstrate the involvement of ultrafast conformational dynamics in allosteric regulation of CIpB, a hexameric AAA+ machine that rescues aggregated proteins. Each subunit of CIpB contains a unique coiled-coil structure, the middle domain (M domain), proposed as a control element that binds the co-chaperone DnaK. Using single-molecule FRET spectroscopy, we probe the M domain during the chaperone cycle and find it to jump on the microsecond time scale between two states, whose structures are determined. The M-domain jumps are much faster than the overall activity of CIpB, making it an effectively continuous, tunable switch. Indeed, a series of allosteric interactions are found to modulate the dynamics, including binding of nucleotides, DnaK and protein substrates. This mode of dynamic control enables fast cellular adaptation and may be a general mechanism for the regulation of cellular machineries.

Original language	English
Article number	1438
Number of pages	12
Journal	Nature Communications
Volume	10
Issue number	1
DOIs	https://doi.org/10.1038/s41467-019-09474-6
Publication status	Published Online - 29 Mar 2019

Funding

G.H. is funded by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 742637). P.G. is funded by the Swiss National Science Foundation (Grant 31003A_156948). The PhD research of H.M. is supported by Planning & Budgeting Committee of the Council of Higher Education of Israel. M.I. is the recipient of an EMBO Long-Term Fellowship (ALTF 317–2018), Y.B. is the incumbent of Beatrice Barton Research Fellowship, and G.H. is the incumbent of the Hilda Pomeraniec Memorial Professorial Chair. We thank Dr. Hagen Hofmann from the Weizmann Institute of Science for reading and commenting on the manuscript, and Prof. Francis Tsai from Baylor College of Medicine, Houston, for discussion and advice at early stages of this work.

All Science Journal Classification (ASJC) codes

General Chemistry
General Biochemistry,Genetics and Molecular Biology
General Physics and Astronomy

Access to Document

10.1038/s41467-019-09474-6Licence: Other

gh_NatCommun_TunableMicrosecondDynamics_PV2019.pdfFinal published version, 1.52 MBLicence: CC BY

Cite this

@article{ff3b9afab6ae4fdb8ccd380e6b94e6cc,

title = "Tunable microsecond dynamics of an allosteric switch regulate the activity of a AAA+ disaggregation machine",

abstract = "Large protein machines are tightly regulated through allosteric communication channels. Here we demonstrate the involvement of ultrafast conformational dynamics in allosteric regulation of CIpB, a hexameric AAA+ machine that rescues aggregated proteins. Each subunit of CIpB contains a unique coiled-coil structure, the middle domain (M domain), proposed as a control element that binds the co-chaperone DnaK. Using single-molecule FRET spectroscopy, we probe the M domain during the chaperone cycle and find it to jump on the microsecond time scale between two states, whose structures are determined. The M-domain jumps are much faster than the overall activity of CIpB, making it an effectively continuous, tunable switch. Indeed, a series of allosteric interactions are found to modulate the dynamics, including binding of nucleotides, DnaK and protein substrates. This mode of dynamic control enables fast cellular adaptation and may be a general mechanism for the regulation of cellular machineries.",

author = "Hisham Mazal and Marija Iljina and Yoav Barak and Nadav Elad and Rina Rosenzweig and Pierre Goloubinoff and Inbal Riven and Gilad Haran",

year = "2019",

month = mar,

day = "29",

doi = "10.1038/s41467-019-09474-6",

language = "English",

volume = "10",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "Nature Publishing Group",

number = "1",

}

TY - JOUR

T1 - Tunable microsecond dynamics of an allosteric switch regulate the activity of a AAA+ disaggregation machine

AU - Mazal, Hisham

AU - Iljina, Marija

AU - Barak, Yoav

AU - Elad, Nadav

AU - Rosenzweig, Rina

AU - Goloubinoff, Pierre

AU - Riven, Inbal

AU - Haran, Gilad

PY - 2019/3/29

Y1 - 2019/3/29

N2 - Large protein machines are tightly regulated through allosteric communication channels. Here we demonstrate the involvement of ultrafast conformational dynamics in allosteric regulation of CIpB, a hexameric AAA+ machine that rescues aggregated proteins. Each subunit of CIpB contains a unique coiled-coil structure, the middle domain (M domain), proposed as a control element that binds the co-chaperone DnaK. Using single-molecule FRET spectroscopy, we probe the M domain during the chaperone cycle and find it to jump on the microsecond time scale between two states, whose structures are determined. The M-domain jumps are much faster than the overall activity of CIpB, making it an effectively continuous, tunable switch. Indeed, a series of allosteric interactions are found to modulate the dynamics, including binding of nucleotides, DnaK and protein substrates. This mode of dynamic control enables fast cellular adaptation and may be a general mechanism for the regulation of cellular machineries.

AB - Large protein machines are tightly regulated through allosteric communication channels. Here we demonstrate the involvement of ultrafast conformational dynamics in allosteric regulation of CIpB, a hexameric AAA+ machine that rescues aggregated proteins. Each subunit of CIpB contains a unique coiled-coil structure, the middle domain (M domain), proposed as a control element that binds the co-chaperone DnaK. Using single-molecule FRET spectroscopy, we probe the M domain during the chaperone cycle and find it to jump on the microsecond time scale between two states, whose structures are determined. The M-domain jumps are much faster than the overall activity of CIpB, making it an effectively continuous, tunable switch. Indeed, a series of allosteric interactions are found to modulate the dynamics, including binding of nucleotides, DnaK and protein substrates. This mode of dynamic control enables fast cellular adaptation and may be a general mechanism for the regulation of cellular machineries.

UR - http://www.scopus.com/inward/record.url?scp=85063740365&partnerID=8YFLogxK

U2 - 10.1038/s41467-019-09474-6

DO - 10.1038/s41467-019-09474-6

M3 - Article

C2 - 30926805

SN - 2041-1723

VL - 10

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 1438

ER -

Tunable microsecond dynamics of an allosteric switch regulate the activity of a AAA+ disaggregation machine

Abstract

Funding

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this