Structural insights into 5 €2 flap DNA unwinding and incision by the human FAN1 dimer

Qi Zhao; Xiaoyu Xue; Simonne Longerich; Patrick Sung; Yong Xiong

doi:10.1038/ncomms6726

Structural insights into 5 €2 flap DNA unwinding and incision by the human FAN1 dimer

Qi Zhao, Xiaoyu Xue, Simonne Longerich, Patrick Sung, Yong Xiong

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

17 Scopus citations

Abstract

Human FANCD2-associated nuclease 1 (FAN1) is a DNA structure-specific nuclease involved in the processing of DNA interstrand crosslinks (ICLs). FAN1 maintains genomic stability and prevents tissue decline in multiple organs, yet it confers ICL-induced anti-cancer drug resistance in several cancer subtypes. Here we report three crystal structures of human FAN1 in complex with a 5 €2 flap DNA substrate, showing that two FAN1 molecules form a head-to-tail dimer to locate the lesion, orient the DNA and unwind a 5 €2 flap for subsequent incision. Biochemical experiments further validate our model for FAN1 action, as structure-informed mutations that disrupt protein dimerization, substrate orientation or flap unwinding impair the structure-specific nuclease activity. Our work elucidates essential aspects of FAN1-DNA lesion recognition and a unique mechanism of incision. These structural insights shed light on the cellular mechanisms underlying organ degeneration protection and cancer drug resistance mediated by FAN1.

Original language	English (US)
Article number	5726
Journal	Nature communications
Volume	5
DOIs	https://doi.org/10.1038/ncomms6726
State	Published - 2014
Externally published	Yes

ASJC Scopus subject areas

Chemistry(all)
Biochemistry, Genetics and Molecular Biology(all)
Physics and Astronomy(all)

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title = "Structural insights into 5 €2 flap DNA unwinding and incision by the human FAN1 dimer",

abstract = "Human FANCD2-associated nuclease 1 (FAN1) is a DNA structure-specific nuclease involved in the processing of DNA interstrand crosslinks (ICLs). FAN1 maintains genomic stability and prevents tissue decline in multiple organs, yet it confers ICL-induced anti-cancer drug resistance in several cancer subtypes. Here we report three crystal structures of human FAN1 in complex with a 5 €2 flap DNA substrate, showing that two FAN1 molecules form a head-to-tail dimer to locate the lesion, orient the DNA and unwind a 5 €2 flap for subsequent incision. Biochemical experiments further validate our model for FAN1 action, as structure-informed mutations that disrupt protein dimerization, substrate orientation or flap unwinding impair the structure-specific nuclease activity. Our work elucidates essential aspects of FAN1-DNA lesion recognition and a unique mechanism of incision. These structural insights shed light on the cellular mechanisms underlying organ degeneration protection and cancer drug resistance mediated by FAN1.",

author = "Qi Zhao and Xiaoyu Xue and Simonne Longerich and Patrick Sung and Yong Xiong",

note = "Funding Information: We thank the staff at the Advanced Photon Source beamline 24-ID, the National Synchrotron Light Source beamlines X25 and X29A. We are grateful to Josef Jiricny (University of Zurich) for the kind gift of the human FAN1 cDNA and Dr. Valeria Busygina, who purified the first batch of full-length FAN1 in the lab, and Mr. William Elliason (Yale Center of Structural Biology core facility), who provided technical support for the SEC MALLS experiment. This work was supported by an Alex{\textquoteright}s Lemonade Stand Foundation Innovation Award and a March of Dimes Foundation Basil O{\textquoteright}Connor Starter Scholar Research award (Y.X.), and by grants ES015252, ES015632 and CA168635 from the US National Institutes of Health.",

year = "2014",

doi = "10.1038/ncomms6726",

language = "English (US)",

volume = "5",

journal = "Nature Communications",

issn = "2041-1723",

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T1 - Structural insights into 5 €2 flap DNA unwinding and incision by the human FAN1 dimer

AU - Zhao, Qi

AU - Xue, Xiaoyu

AU - Longerich, Simonne

AU - Sung, Patrick

AU - Xiong, Yong

N1 - Funding Information: We thank the staff at the Advanced Photon Source beamline 24-ID, the National Synchrotron Light Source beamlines X25 and X29A. We are grateful to Josef Jiricny (University of Zurich) for the kind gift of the human FAN1 cDNA and Dr. Valeria Busygina, who purified the first batch of full-length FAN1 in the lab, and Mr. William Elliason (Yale Center of Structural Biology core facility), who provided technical support for the SEC MALLS experiment. This work was supported by an Alex’s Lemonade Stand Foundation Innovation Award and a March of Dimes Foundation Basil O’Connor Starter Scholar Research award (Y.X.), and by grants ES015252, ES015632 and CA168635 from the US National Institutes of Health.

PY - 2014

Y1 - 2014

N2 - Human FANCD2-associated nuclease 1 (FAN1) is a DNA structure-specific nuclease involved in the processing of DNA interstrand crosslinks (ICLs). FAN1 maintains genomic stability and prevents tissue decline in multiple organs, yet it confers ICL-induced anti-cancer drug resistance in several cancer subtypes. Here we report three crystal structures of human FAN1 in complex with a 5 €2 flap DNA substrate, showing that two FAN1 molecules form a head-to-tail dimer to locate the lesion, orient the DNA and unwind a 5 €2 flap for subsequent incision. Biochemical experiments further validate our model for FAN1 action, as structure-informed mutations that disrupt protein dimerization, substrate orientation or flap unwinding impair the structure-specific nuclease activity. Our work elucidates essential aspects of FAN1-DNA lesion recognition and a unique mechanism of incision. These structural insights shed light on the cellular mechanisms underlying organ degeneration protection and cancer drug resistance mediated by FAN1.

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Structural insights into 5 €2 flap DNA unwinding and incision by the human FAN1 dimer

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