A minimal threshold of FANCJ helicase activity is required for its response to replication stress or double-strand break repair

Sanjay Kumar Bharti; Joshua A Sommers; Sanket Awate; Marina A Bellani; Irfan Khan; Lynda Bradley; Graeme A King; Yeonee Seol; Venkatasubramanian Vidhyasagar; Yuliang Wu; Takuye Abe; Koji Kobayashi; Kazuo Shin-ya; Hiroyuki Kitao; Marc S Wold; Dana Branzei; Keir C Neuman; Robert M Brosh

doi:10.1093/nar/gky403

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A minimal threshold of FANCJ helicase activity is required for its response to replication stress or double-strand break repair

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A minimal threshold of FANCJ helicase activity is required for its response to replication stress or double-strand break repair

Sanjay Kumar Bharti, Joshua A Sommers, Sanket Awate, Marina A Bellani, Irfan Khan, Lynda Bradley, Graeme A King, Yeonee Seol, Venkatasubramanian Vidhyasagar, Yuliang Wu, …

Nucleic acids research, Vol.46(12), pp.6238-6256

07/06/2018

DOI: 10.1093/nar/gky403

PMCID: PMC6159516

PMID: 29788478

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Abstract

Fanconi Anemia (FA) is characterized by bone marrow failure, congenital abnormalities, and cancer. Of over 20 FA-linked genes, FANCJ uniquely encodes a DNA helicase and mutations are also associated with breast and ovarian cancer. fancj −/− cells are sensitive to DNA interstrand cross-linking (ICL) and replication fork stalling drugs. We delineated the molecular defects of two FA patient-derived FANCJ helicase domain mutations. FANCJ-R707C was compromised in dimerization and helicase processivity, whereas DNA unwinding by FANCJ-H396D was barely detectable. DNA binding and ATP hydrolysis was defective for both FANCJ-R707C and FANCJ-H396D, the latter showing greater reduction. Expression of FANCJ-R707C or FANCJ-H396D in fancj −/− cells failed to rescue cisplatin or mitomycin sensitivity. Live-cell imaging demonstrated a significantly compromised recruitment of FANCJ-R707C to laser-induced DNA damage. However, FANCJ-R707C expressed in fancj-/- cells conferred resistance to the DNA polymerase inhibitor aphidicolin, G-quadruplex ligand telomestatin, or DNA strand-breaker bleomycin, whereas FANCJ-H396D failed. Thus, a minimal threshold of FANCJ catalytic activity is required to overcome replication stress induced by aphidicolin or telomestatin, or to repair bleomycin-induced DNA breakage. These findings have implications for therapeutic strategies relying on DNA cross-link sensitivity or heightened replication stress characteristic of cancer cells.

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Title: Subtitle: A minimal threshold of FANCJ helicase activity is required for its response to replication stress or double-strand break repair
Creators: Sanjay Kumar Bharti - Laboratory of Molecular Gerontology, National Institute on Aging, NIH, NIH Biomedical Research Center, 251 Bayview Blvd, Baltimore, MD 21224, USA
Joshua A Sommers - Laboratory of Molecular Gerontology, National Institute on Aging, NIH, NIH Biomedical Research Center, 251 Bayview Blvd, Baltimore, MD 21224, USA
Sanket Awate - Laboratory of Molecular Gerontology, National Institute on Aging, NIH, NIH Biomedical Research Center, 251 Bayview Blvd, Baltimore, MD 21224, USA
Marina A Bellani - Laboratory of Molecular Gerontology, National Institute on Aging, NIH, NIH Biomedical Research Center, 251 Bayview Blvd, Baltimore, MD 21224, USA
Irfan Khan - Laboratory of Molecular Gerontology, National Institute on Aging, NIH, NIH Biomedical Research Center, 251 Bayview Blvd, Baltimore, MD 21224, USA
Lynda Bradley - Laboratory of Single Molecule Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
Graeme A King - Laboratory of Single Molecule Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
Yeonee Seol - Laboratory of Single Molecule Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
Venkatasubramanian Vidhyasagar - Department of Biochemistry, University of Saskatchewan, Health Sciences Building, 107 Wiggins Road, Saskatoon, Saskatchewan S7N 5E5, Canada
Yuliang Wu - Department of Biochemistry, University of Saskatchewan, Health Sciences Building, 107 Wiggins Road, Saskatoon, Saskatchewan S7N 5E5, Canada
Takuye Abe - IFOM, the FIRC Institute for Molecular Oncology Foundation, Milan, Italy
Koji Kobayashi - IFOM, the FIRC Institute for Molecular Oncology Foundation, Milan, Italy
Kazuo Shin-ya - Department of Life Science and Biotechnology Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST) 2-4-7 Aomi, Koto-ku, Tokyo 135-0064, Japan
Hiroyuki Kitao - Department of Molecular Cancer Biology, Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan
Marc S Wold - Department of Biochemistry, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA
Dana Branzei - IFOM, the FIRC Institute for Molecular Oncology Foundation, Milan, Italy
Keir C Neuman - Laboratory of Single Molecule Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
Robert M Brosh - Laboratory of Molecular Gerontology, National Institute on Aging, NIH, NIH Biomedical Research Center, 251 Bayview Blvd, Baltimore, MD 21224, USA
Resource Type: Journal article
Publication Details: Nucleic acids research, Vol.46(12), pp.6238-6256
Publisher: Oxford University Press
DOI: 10.1093/nar/gky403
PMID: 29788478
PMCID: PMC6159516
ISSN: 0305-1048
eISSN: 1362-4962
Grant note: 14171 / ; ; ;
Language: English
Date published: 07/06/2018
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984025398502771

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