Reverse-chaperoning activity of an AAA+ protein

Cheng Liu; Mary C. McKinney; Yi Hsing Chen; Tyler M. Earnest; Xinghua Shi; Li Jung Lin; Yoshizumi Ishino; Karin Dahmen; Isaac K.O. Cann; Taekjip Ha

doi:10.1016/j.bpj.2011.01.057

Reverse-chaperoning activity of an AAA+ protein

Cheng Liu, Mary C. McKinney, Yi Hsing Chen, Tyler M. Earnest, Xinghua Shi, Li Jung Lin, Yoshizumi Ishino, Karin Dahmen, Isaac K.O. Cann, Taekjip Ha

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

Abstract

Speed and processivity of replicative DNA polymerases can be enhanced via coupling to a sliding clamp. Due to the closed ring shape of the clamp, a clamp loader protein, belonging to the AAA+ class of ATPases, needs to open the ring-shaped clamp before loading it to DNA. Here, we developed real-time fluorescence assays to study the clamp (PCNA) and the clamp loader (RFC) from the mesophilic archaeon Methanosarcina acetivorans. Unexpectedly, we discovered that RFC can assemble a PCNA ring from monomers in solution. A motion-based DNA polymerization assay showed that the PCNA assembled by RFC is functional. This PCNA assembly activity required the ATP-bound conformation of RFC. Our work demonstrates a reverse-chaperoning activity for an AAA+ protein that can act as a template for the assembly of another protein complex.

Original language	English (US)
Pages (from-to)	1344-1352
Number of pages	9
Journal	Biophysical journal
Volume	100
Issue number	5
DOIs	https://doi.org/10.1016/j.bpj.2011.01.057
State	Published - Mar 2 2011

ASJC Scopus subject areas

Biophysics

Online availability

10.1016/j.bpj.2011.01.057

Library availability

Discover UIUC Full Text

Cite this

@article{7ce4a3d830df486f893888b2b034de5a,

title = "Reverse-chaperoning activity of an AAA+ protein",

abstract = "Speed and processivity of replicative DNA polymerases can be enhanced via coupling to a sliding clamp. Due to the closed ring shape of the clamp, a clamp loader protein, belonging to the AAA+ class of ATPases, needs to open the ring-shaped clamp before loading it to DNA. Here, we developed real-time fluorescence assays to study the clamp (PCNA) and the clamp loader (RFC) from the mesophilic archaeon Methanosarcina acetivorans. Unexpectedly, we discovered that RFC can assemble a PCNA ring from monomers in solution. A motion-based DNA polymerization assay showed that the PCNA assembled by RFC is functional. This PCNA assembly activity required the ATP-bound conformation of RFC. Our work demonstrates a reverse-chaperoning activity for an AAA+ protein that can act as a template for the assembly of another protein complex.",

author = "Cheng Liu and McKinney, {Mary C.} and Chen, {Yi Hsing} and Earnest, {Tyler M.} and Xinghua Shi and Lin, {Li Jung} and Yoshizumi Ishino and Karin Dahmen and Cann, {Isaac K.O.} and Taekjip Ha",

note = "Funding Information: This work was supported by the National Science Foundation (grant Nos. MCB-0238451, PHY-0822613, and PHY-0646550), the National Institutes of Health (grant No. GM065367), and the Human Frontiers Science Program. M.C.M. was supported by Agricultural Genome Sciences and Public Policy training grant No. 2001-52100-11527. ",

year = "2011",

month = mar,

day = "2",

doi = "10.1016/j.bpj.2011.01.057",

language = "English (US)",

volume = "100",

pages = "1344--1352",

journal = "Biophysical journal",

issn = "0006-3495",

publisher = "Elsevier B.V.",

number = "5",

}

TY - JOUR

T1 - Reverse-chaperoning activity of an AAA+ protein

AU - Liu, Cheng

AU - McKinney, Mary C.

AU - Chen, Yi Hsing

AU - Earnest, Tyler M.

AU - Shi, Xinghua

AU - Lin, Li Jung

AU - Ishino, Yoshizumi

AU - Dahmen, Karin

AU - Cann, Isaac K.O.

AU - Ha, Taekjip

N1 - Funding Information: This work was supported by the National Science Foundation (grant Nos. MCB-0238451, PHY-0822613, and PHY-0646550), the National Institutes of Health (grant No. GM065367), and the Human Frontiers Science Program. M.C.M. was supported by Agricultural Genome Sciences and Public Policy training grant No. 2001-52100-11527.

PY - 2011/3/2

Y1 - 2011/3/2

N2 - Speed and processivity of replicative DNA polymerases can be enhanced via coupling to a sliding clamp. Due to the closed ring shape of the clamp, a clamp loader protein, belonging to the AAA+ class of ATPases, needs to open the ring-shaped clamp before loading it to DNA. Here, we developed real-time fluorescence assays to study the clamp (PCNA) and the clamp loader (RFC) from the mesophilic archaeon Methanosarcina acetivorans. Unexpectedly, we discovered that RFC can assemble a PCNA ring from monomers in solution. A motion-based DNA polymerization assay showed that the PCNA assembled by RFC is functional. This PCNA assembly activity required the ATP-bound conformation of RFC. Our work demonstrates a reverse-chaperoning activity for an AAA+ protein that can act as a template for the assembly of another protein complex.

AB - Speed and processivity of replicative DNA polymerases can be enhanced via coupling to a sliding clamp. Due to the closed ring shape of the clamp, a clamp loader protein, belonging to the AAA+ class of ATPases, needs to open the ring-shaped clamp before loading it to DNA. Here, we developed real-time fluorescence assays to study the clamp (PCNA) and the clamp loader (RFC) from the mesophilic archaeon Methanosarcina acetivorans. Unexpectedly, we discovered that RFC can assemble a PCNA ring from monomers in solution. A motion-based DNA polymerization assay showed that the PCNA assembled by RFC is functional. This PCNA assembly activity required the ATP-bound conformation of RFC. Our work demonstrates a reverse-chaperoning activity for an AAA+ protein that can act as a template for the assembly of another protein complex.

UR - http://www.scopus.com/inward/record.url?scp=79953837287&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79953837287&partnerID=8YFLogxK

U2 - 10.1016/j.bpj.2011.01.057

DO - 10.1016/j.bpj.2011.01.057

M3 - Article

C2 - 21354408

AN - SCOPUS:79953837287

SN - 0006-3495

VL - 100

SP - 1344

EP - 1352

JO - Biophysical journal

JF - Biophysical journal

IS - 5

ER -

Reverse-chaperoning activity of an AAA+ protein

Abstract

ASJC Scopus subject areas

Online availability

Library availability

Related links

Fingerprint

Cite this