Nucleotide-induced conformational changes in an isolated Escherichia coli DNA polymerase III clamp loader subunit

Marjetka Podobnik; Tanya F. Weitze; Mike O'Donnell; John Kuriyan

doi:10.1016/S0969-2126(03)00027-3

Nucleotide-induced conformational changes in an isolated Escherichia coli DNA polymerase III clamp loader subunit

Marjetka Podobnik, Tanya F. Weitze, Mike O'Donnell, John Kuriyan

Pharmacology

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

28 Scopus citations

Abstract

Sliding clamps are loaded onto DNA by ATP-driven clamp loader complexes. The structure of the E. coli clamp loader in a nucleotide-free state has been determined previously. We now report crystal structures of a truncated form of the isolated γ-ATPase subunit, γ^1-243, of the E. coli clamp loader, in nucleotide-free and bound forms. The γ subunit adopts a defined conformation when empty, in which the nucleotide binding site is blocked. The binding of either ATPγS or ADP, which are shown to bind with equal affinity to γ^1-243, induces a change in the relative orientation of the two domains such that nucleotides can be accommodated. This change would break one of the γ:γ interfaces seen in the empty clamp loader complex, and may represent one step in the activation process.

Original language	English (US)
Pages (from-to)	253-263
Number of pages	11
Journal	Structure
Volume	11
Issue number	3
DOIs	https://doi.org/10.1016/S0969-2126(03)00027-3
State	Published - Mar 1 2003

Bibliographical note

Funding Information:
We thank David Jeruzalmi, Gregory D. Bowman, Holger Sondermann, and Bhushan Nagar for valuable assistance and discussions. We thank the staff at the 5.0.2 and 8.3.1 beamlines at the Advanced Light Source, Berkeley for assistance with synchrotron data collection. This work was supported by NIH grants GM45547 (J.K.) and GM38839 (M.O.D).

Keywords

AAA+ ATPase
Clamp loader
DNA polymerase III
Replication factor C (RFC)
γ subunit

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abstract = "Sliding clamps are loaded onto DNA by ATP-driven clamp loader complexes. The structure of the E. coli clamp loader in a nucleotide-free state has been determined previously. We now report crystal structures of a truncated form of the isolated γ-ATPase subunit, γ1-243, of the E. coli clamp loader, in nucleotide-free and bound forms. The γ subunit adopts a defined conformation when empty, in which the nucleotide binding site is blocked. The binding of either ATPγS or ADP, which are shown to bind with equal affinity to γ1-243, induces a change in the relative orientation of the two domains such that nucleotides can be accommodated. This change would break one of the γ:γ interfaces seen in the empty clamp loader complex, and may represent one step in the activation process.",

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author = "Marjetka Podobnik and Weitze, {Tanya F.} and Mike O'Donnell and John Kuriyan",

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AU - Weitze, Tanya F.

AU - O'Donnell, Mike

AU - Kuriyan, John

N1 - Funding Information: We thank David Jeruzalmi, Gregory D. Bowman, Holger Sondermann, and Bhushan Nagar for valuable assistance and discussions. We thank the staff at the 5.0.2 and 8.3.1 beamlines at the Advanced Light Source, Berkeley for assistance with synchrotron data collection. This work was supported by NIH grants GM45547 (J.K.) and GM38839 (M.O.D).

PY - 2003/3/1

Y1 - 2003/3/1

N2 - Sliding clamps are loaded onto DNA by ATP-driven clamp loader complexes. The structure of the E. coli clamp loader in a nucleotide-free state has been determined previously. We now report crystal structures of a truncated form of the isolated γ-ATPase subunit, γ1-243, of the E. coli clamp loader, in nucleotide-free and bound forms. The γ subunit adopts a defined conformation when empty, in which the nucleotide binding site is blocked. The binding of either ATPγS or ADP, which are shown to bind with equal affinity to γ1-243, induces a change in the relative orientation of the two domains such that nucleotides can be accommodated. This change would break one of the γ:γ interfaces seen in the empty clamp loader complex, and may represent one step in the activation process.

AB - Sliding clamps are loaded onto DNA by ATP-driven clamp loader complexes. The structure of the E. coli clamp loader in a nucleotide-free state has been determined previously. We now report crystal structures of a truncated form of the isolated γ-ATPase subunit, γ1-243, of the E. coli clamp loader, in nucleotide-free and bound forms. The γ subunit adopts a defined conformation when empty, in which the nucleotide binding site is blocked. The binding of either ATPγS or ADP, which are shown to bind with equal affinity to γ1-243, induces a change in the relative orientation of the two domains such that nucleotides can be accommodated. This change would break one of the γ:γ interfaces seen in the empty clamp loader complex, and may represent one step in the activation process.

KW - AAA+ ATPase

KW - Clamp loader

KW - DNA polymerase III

KW - Replication factor C (RFC)

KW - γ subunit

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Nucleotide-induced conformational changes in an isolated Escherichia coli DNA polymerase III clamp loader subunit

Abstract

Bibliographical note

Keywords

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