A Flexible Interface between DNA Ligase and PCNA Supports Conformational Switching and Efficient Ligation of DNA

John M. Pascal; Oleg V. Tsodikov; Greg L. Hura; Wei Song; Elizabeth A. Cotner; Scott Classen; Alan E. Tomkinson; John A. Tainer; Tom Ellenberger

doi:10.1016/j.molcel.2006.08.015

A Flexible Interface between DNA Ligase and PCNA Supports Conformational Switching and Efficient Ligation of DNA

John M. Pascal, Oleg V. Tsodikov, Greg L. Hura, Wei Song, Elizabeth A. Cotner, Scott Classen, Alan E. Tomkinson, John A. Tainer, Tom Ellenberger

Producción científica: Article › revisión exhaustiva

146 Citas (Scopus)

Resumen

DNA sliding clamps encircle DNA and provide binding sites for many DNA-processing enzymes. However, it is largely unknown how sliding clamps like proliferating cell nuclear antigen (PCNA) coordinate multistep DNA transactions. We have determined structures of Sulfolobus solfataricus DNA ligase and heterotrimeric PCNA separately by X-ray diffraction and in complex by small-angle X-ray scattering (SAXS). Three distinct PCNA subunits assemble into a protein ring resembling the homotrimeric PCNA of humans but with three unique protein-binding sites. In the absence of nicked DNA, the Sulfolobus solfataricus DNA ligase has an open, extended conformation. When complexed with heterotrimeric PCNA, the DNA ligase binds to the PCNA3 subunit and ligase retains an open, extended conformation. A closed, ring-shaped conformation of ligase catalyzes a DNA end-joining reaction that is strongly stimulated by PCNA. This open-to-closed switch in the conformation of DNA ligase is accommodated by a malleable interface with PCNA that serves as an efficient platform for DNA ligation.

Idioma original	English
Páginas (desde-hasta)	279-291
Número de páginas	13
Publicación	Molecular Cell
Volumen	24
N.º	2
DOI	https://doi.org/10.1016/j.molcel.2006.08.015
Estado	Published - oct 20 2006

Nota bibliográfica

Funding Information:
We thank the beamline staff at ALS-SIBYLS for help with experiments, and we thank members of our laboratories for thoughtful discussion. This work was supported by a grant from the National Institute of General Medical Sciences (GM52504 to T.E.), by National Cancer Institute grants (CA081967 to J.A.T.), and by program P01 CA92584 (to J.A.T., A.E.T., and T.E.). We thank the U.S. Department of Energy for support of the SIBYLS beamline and of SAXS experiments under contract number DE-AC02-05CH11231.

Financiación

We thank the beamline staff at ALS-SIBYLS for help with experiments, and we thank members of our laboratories for thoughtful discussion. This work was supported by a grant from the National Institute of General Medical Sciences (GM52504 to T.E.), by National Cancer Institute grants (CA081967 to J.A.T.), and by program P01 CA92584 (to J.A.T., A.E.T., and T.E.). We thank the U.S. Department of Energy for support of the SIBYLS beamline and of SAXS experiments under contract number DE-AC02-05CH11231.

Financiadores	Número del financiador
Michigan State University-U.S. Department of Energy (MSU-DOE) Plant Research Laboratory	DE-AC02-05CH11231
National Childhood Cancer Registry – National Cancer Institute	CA081967, P01CA092584
National Institute of General Medical Sciences	GM52504

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1016/j.molcel.2006.08.015

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title = "A Flexible Interface between DNA Ligase and PCNA Supports Conformational Switching and Efficient Ligation of DNA",

abstract = "DNA sliding clamps encircle DNA and provide binding sites for many DNA-processing enzymes. However, it is largely unknown how sliding clamps like proliferating cell nuclear antigen (PCNA) coordinate multistep DNA transactions. We have determined structures of Sulfolobus solfataricus DNA ligase and heterotrimeric PCNA separately by X-ray diffraction and in complex by small-angle X-ray scattering (SAXS). Three distinct PCNA subunits assemble into a protein ring resembling the homotrimeric PCNA of humans but with three unique protein-binding sites. In the absence of nicked DNA, the Sulfolobus solfataricus DNA ligase has an open, extended conformation. When complexed with heterotrimeric PCNA, the DNA ligase binds to the PCNA3 subunit and ligase retains an open, extended conformation. A closed, ring-shaped conformation of ligase catalyzes a DNA end-joining reaction that is strongly stimulated by PCNA. This open-to-closed switch in the conformation of DNA ligase is accommodated by a malleable interface with PCNA that serves as an efficient platform for DNA ligation.",

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author = "Pascal, \{John M.\} and Tsodikov, \{Oleg V.\} and Hura, \{Greg L.\} and Wei Song and Cotner, \{Elizabeth A.\} and Scott Classen and Tomkinson, \{Alan E.\} and Tainer, \{John A.\} and Tom Ellenberger",

note = "Funding Information: We thank the beamline staff at ALS-SIBYLS for help with experiments, and we thank members of our laboratories for thoughtful discussion. This work was supported by a grant from the National Institute of General Medical Sciences (GM52504 to T.E.), by National Cancer Institute grants (CA081967 to J.A.T.), and by program P01 CA92584 (to J.A.T., A.E.T., and T.E.). We thank the U.S. Department of Energy for support of the SIBYLS beamline and of SAXS experiments under contract number DE-AC02-05CH11231. ",

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AU - Pascal, John M.

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AU - Song, Wei

AU - Cotner, Elizabeth A.

AU - Classen, Scott

AU - Tomkinson, Alan E.

AU - Tainer, John A.

AU - Ellenberger, Tom

N1 - Funding Information: We thank the beamline staff at ALS-SIBYLS for help with experiments, and we thank members of our laboratories for thoughtful discussion. This work was supported by a grant from the National Institute of General Medical Sciences (GM52504 to T.E.), by National Cancer Institute grants (CA081967 to J.A.T.), and by program P01 CA92584 (to J.A.T., A.E.T., and T.E.). We thank the U.S. Department of Energy for support of the SIBYLS beamline and of SAXS experiments under contract number DE-AC02-05CH11231.

PY - 2006/10/20

Y1 - 2006/10/20

N2 - DNA sliding clamps encircle DNA and provide binding sites for many DNA-processing enzymes. However, it is largely unknown how sliding clamps like proliferating cell nuclear antigen (PCNA) coordinate multistep DNA transactions. We have determined structures of Sulfolobus solfataricus DNA ligase and heterotrimeric PCNA separately by X-ray diffraction and in complex by small-angle X-ray scattering (SAXS). Three distinct PCNA subunits assemble into a protein ring resembling the homotrimeric PCNA of humans but with three unique protein-binding sites. In the absence of nicked DNA, the Sulfolobus solfataricus DNA ligase has an open, extended conformation. When complexed with heterotrimeric PCNA, the DNA ligase binds to the PCNA3 subunit and ligase retains an open, extended conformation. A closed, ring-shaped conformation of ligase catalyzes a DNA end-joining reaction that is strongly stimulated by PCNA. This open-to-closed switch in the conformation of DNA ligase is accommodated by a malleable interface with PCNA that serves as an efficient platform for DNA ligation.

AB - DNA sliding clamps encircle DNA and provide binding sites for many DNA-processing enzymes. However, it is largely unknown how sliding clamps like proliferating cell nuclear antigen (PCNA) coordinate multistep DNA transactions. We have determined structures of Sulfolobus solfataricus DNA ligase and heterotrimeric PCNA separately by X-ray diffraction and in complex by small-angle X-ray scattering (SAXS). Three distinct PCNA subunits assemble into a protein ring resembling the homotrimeric PCNA of humans but with three unique protein-binding sites. In the absence of nicked DNA, the Sulfolobus solfataricus DNA ligase has an open, extended conformation. When complexed with heterotrimeric PCNA, the DNA ligase binds to the PCNA3 subunit and ligase retains an open, extended conformation. A closed, ring-shaped conformation of ligase catalyzes a DNA end-joining reaction that is strongly stimulated by PCNA. This open-to-closed switch in the conformation of DNA ligase is accommodated by a malleable interface with PCNA that serves as an efficient platform for DNA ligation.

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A Flexible Interface between DNA Ligase and PCNA Supports Conformational Switching and Efficient Ligation of DNA

Resumen

Nota bibliográfica

Financiación

ASJC Scopus subject areas

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