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 journalArticle

5 Citations (Scopus)

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 languageEnglish
Pages (from-to)1344-1352
Number of pages9
JournalBiophysical Journal
Volume100
Issue number5
DOIs
Publication statusPublished - Mar 2 2011

Fingerprint

Proliferating Cell Nuclear Antigen
Proteins
Methanosarcina
DNA
Archaea
DNA-Directed DNA Polymerase
Polymerization
Adenosine Triphosphatases
Adenosine Triphosphate
Fluorescence

All Science Journal Classification (ASJC) codes

  • Biophysics

Cite this

Liu, C., McKinney, M. C., Chen, Y. H., Earnest, T. M., Shi, X., Lin, L. J., ... Ha, T. (2011). Reverse-chaperoning activity of an AAA+ protein. Biophysical Journal, 100(5), 1344-1352. https://doi.org/10.1016/j.bpj.2011.01.057

Reverse-chaperoning activity of an AAA+ protein. / Liu, Cheng; McKinney, Mary C.; Chen, Yi Hsing; Earnest, Tyler M.; Shi, Xinghua; Lin, Li Jung; Ishino, Yoshizumi; Dahmen, Karin; Cann, Isaac K.O.; Ha, Taekjip.

In: Biophysical Journal, Vol. 100, No. 5, 02.03.2011, p. 1344-1352.

Research output: Contribution to journalArticle

Liu, C, McKinney, MC, Chen, YH, Earnest, TM, Shi, X, Lin, LJ, Ishino, Y, Dahmen, K, Cann, IKO & Ha, T 2011, 'Reverse-chaperoning activity of an AAA+ protein', Biophysical Journal, vol. 100, no. 5, pp. 1344-1352. https://doi.org/10.1016/j.bpj.2011.01.057
Liu C, McKinney MC, Chen YH, Earnest TM, Shi X, Lin LJ et al. Reverse-chaperoning activity of an AAA+ protein. Biophysical Journal. 2011 Mar 2;100(5):1344-1352. https://doi.org/10.1016/j.bpj.2011.01.057
Liu, Cheng ; McKinney, Mary C. ; Chen, Yi Hsing ; Earnest, Tyler M. ; Shi, Xinghua ; Lin, Li Jung ; Ishino, Yoshizumi ; Dahmen, Karin ; Cann, Isaac K.O. ; Ha, Taekjip. / Reverse-chaperoning activity of an AAA+ protein. In: Biophysical Journal. 2011 ; Vol. 100, No. 5. pp. 1344-1352.
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