Structures of Minimal Catalytic Fragments of Topoisomerase V Reveals Conformational Changes Relevant for DNA Binding

Rakhi Rajan; Bhupesh Taneja; Alfonso Mondragón

doi:10.1016/j.str.2010.03.006

Structures of Minimal Catalytic Fragments of Topoisomerase V Reveals Conformational Changes Relevant for DNA Binding

Rakhi Rajan, Bhupesh Taneja, Alfonso Mondragón^*

^*Corresponding author for this work

Molecular Biosciences

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

9 Scopus citations

Abstract

Topoisomerase V is an archaeal type I topoisomerase that is unique among topoisomerases due to presence of both topoisomerase and DNA repair activities in the same protein. It is organized as an N-terminal topoisomerase domain followed by 24 tandem helix-hairpin-helix (HhH) motifs. Structural studies have shown that the active site is buried by the (HhH) motifs. Here we show that the N-terminal domain can relax DNA in the absence of any HhH motifs and that the HhH motifs are required for stable protein-DNA complex formation. Crystal structures of various topoisomerase V fragments show changes in the relative orientation of the domains mediated by a long bent linker helix, and these movements are essential for the DNA to enter the active site. Phosphate ions bound to the protein near the active site helped model DNA in the topoisomerase domain and show how topoisomerase V may interact with DNA.

Original language	English (US)
Pages (from-to)	829-838
Number of pages	10
Journal	Structure
Volume	18
Issue number	7
DOIs	https://doi.org/10.1016/j.str.2010.03.006
State	Published - Jul 2010

Funding

We acknowledge staff and instrumentation support from the Keck Biophysics Facility and the Center for Structural Biology at Northwestern University and from Dupont Northwestern Dow (DND) and Life Science Collaborative Access Team (LS-CAT) stations at the Advanced Photon Source (APS) at Argonne National Laboratory. Support from the R.H. Lurie Comprehensive Cancer Center of Northwestern University to the Structural Biology Facility is also acknowledged. DND-CAT is supported by Dupont, DOW, and the National Science Foundation. LS-CAT was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor. Use of the APS is supported by the Department of Energy. Research was supported by National Institutes of Health grant GM51350 (to A.M.).

Keywords

DNA
Proteins

ASJC Scopus subject areas

Molecular Biology
Structural Biology

Access to Document

10.1016/j.str.2010.03.006

Cite this

@article{188338620cab4eb18b02ccf3347a4175,

title = "Structures of Minimal Catalytic Fragments of Topoisomerase V Reveals Conformational Changes Relevant for DNA Binding",

abstract = "Topoisomerase V is an archaeal type I topoisomerase that is unique among topoisomerases due to presence of both topoisomerase and DNA repair activities in the same protein. It is organized as an N-terminal topoisomerase domain followed by 24 tandem helix-hairpin-helix (HhH) motifs. Structural studies have shown that the active site is buried by the (HhH) motifs. Here we show that the N-terminal domain can relax DNA in the absence of any HhH motifs and that the HhH motifs are required for stable protein-DNA complex formation. Crystal structures of various topoisomerase V fragments show changes in the relative orientation of the domains mediated by a long bent linker helix, and these movements are essential for the DNA to enter the active site. Phosphate ions bound to the protein near the active site helped model DNA in the topoisomerase domain and show how topoisomerase V may interact with DNA.",

keywords = "DNA, Proteins",

author = "Rakhi Rajan and Bhupesh Taneja and Alfonso Mondrag{\'o}n",

note = "Funding Information: We acknowledge staff and instrumentation support from the Keck Biophysics Facility and the Center for Structural Biology at Northwestern University and from Dupont Northwestern Dow (DND) and Life Science Collaborative Access Team (LS-CAT) stations at the Advanced Photon Source (APS) at Argonne National Laboratory. Support from the R.H. Lurie Comprehensive Cancer Center of Northwestern University to the Structural Biology Facility is also acknowledged. DND-CAT is supported by Dupont, DOW, and the National Science Foundation. LS-CAT was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor. Use of the APS is supported by the Department of Energy. Research was supported by National Institutes of Health grant GM51350 (to A.M.). ",

year = "2010",

month = jul,

doi = "10.1016/j.str.2010.03.006",

language = "English (US)",

volume = "18",

pages = "829--838",

journal = "Structure",

issn = "0969-2126",

publisher = "Cell Press",

number = "7",

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AU - Rajan, Rakhi

AU - Taneja, Bhupesh

AU - Mondragón, Alfonso

N1 - Funding Information: We acknowledge staff and instrumentation support from the Keck Biophysics Facility and the Center for Structural Biology at Northwestern University and from Dupont Northwestern Dow (DND) and Life Science Collaborative Access Team (LS-CAT) stations at the Advanced Photon Source (APS) at Argonne National Laboratory. Support from the R.H. Lurie Comprehensive Cancer Center of Northwestern University to the Structural Biology Facility is also acknowledged. DND-CAT is supported by Dupont, DOW, and the National Science Foundation. LS-CAT was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor. Use of the APS is supported by the Department of Energy. Research was supported by National Institutes of Health grant GM51350 (to A.M.).

PY - 2010/7

Y1 - 2010/7

N2 - Topoisomerase V is an archaeal type I topoisomerase that is unique among topoisomerases due to presence of both topoisomerase and DNA repair activities in the same protein. It is organized as an N-terminal topoisomerase domain followed by 24 tandem helix-hairpin-helix (HhH) motifs. Structural studies have shown that the active site is buried by the (HhH) motifs. Here we show that the N-terminal domain can relax DNA in the absence of any HhH motifs and that the HhH motifs are required for stable protein-DNA complex formation. Crystal structures of various topoisomerase V fragments show changes in the relative orientation of the domains mediated by a long bent linker helix, and these movements are essential for the DNA to enter the active site. Phosphate ions bound to the protein near the active site helped model DNA in the topoisomerase domain and show how topoisomerase V may interact with DNA.

AB - Topoisomerase V is an archaeal type I topoisomerase that is unique among topoisomerases due to presence of both topoisomerase and DNA repair activities in the same protein. It is organized as an N-terminal topoisomerase domain followed by 24 tandem helix-hairpin-helix (HhH) motifs. Structural studies have shown that the active site is buried by the (HhH) motifs. Here we show that the N-terminal domain can relax DNA in the absence of any HhH motifs and that the HhH motifs are required for stable protein-DNA complex formation. Crystal structures of various topoisomerase V fragments show changes in the relative orientation of the domains mediated by a long bent linker helix, and these movements are essential for the DNA to enter the active site. Phosphate ions bound to the protein near the active site helped model DNA in the topoisomerase domain and show how topoisomerase V may interact with DNA.

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Structures of Minimal Catalytic Fragments of Topoisomerase V Reveals Conformational Changes Relevant for DNA Binding

Abstract

Funding

Keywords

ASJC Scopus subject areas

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Fingerprint

Crystal Structure of N-terminal 44 kDa fragment of topoisomerase V in the presence of guanidium hydrochloride

Crystal structure of an N-terminal 44 kDA fragment of topoisomerase V in the presence of dioxane

Structure of topoisomerase domain of topoisomerase V protein

Cite this

Structures of Minimal Catalytic Fragments of Topoisomerase V Reveals Conformational Changes Relevant for DNA Binding

Abstract

Funding

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Datasets

Crystal Structure of N-terminal 44 kDa fragment of topoisomerase V in the presence of guanidium hydrochloride

Crystal structure of an N-terminal 44 kDA fragment of topoisomerase V in the presence of dioxane

Structure of topoisomerase domain of topoisomerase V protein

Cite this