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

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)...

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Autores principales: Bharti, Sanjay Kumar, Sommers, Joshua A, Awate, Sanket, Bellani, Marina A, Khan, Irfan, Bradley, Lynda, King, Graeme A, Seol, Yeonee, Vidhyasagar, Venkatasubramanian, Wu, Yuliang, Abe, Takuye, Kobayashi, Koji, Shin-ya, Kazuo, Kitao, Hiroyuki, Wold, Marc S, Branzei, Dana, Neuman, Keir C, Brosh, Robert M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6159516/
https://www.ncbi.nlm.nih.gov/pubmed/29788478
http://dx.doi.org/10.1093/nar/gky403
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author Bharti, Sanjay Kumar
Sommers, Joshua A
Awate, Sanket
Bellani, Marina A
Khan, Irfan
Bradley, Lynda
King, Graeme A
Seol, Yeonee
Vidhyasagar, Venkatasubramanian
Wu, Yuliang
Abe, Takuye
Kobayashi, Koji
Shin-ya, Kazuo
Kitao, Hiroyuki
Wold, Marc S
Branzei, Dana
Neuman, Keir C
Brosh, Robert M
author_facet Bharti, Sanjay Kumar
Sommers, Joshua A
Awate, Sanket
Bellani, Marina A
Khan, Irfan
Bradley, Lynda
King, Graeme A
Seol, Yeonee
Vidhyasagar, Venkatasubramanian
Wu, Yuliang
Abe, Takuye
Kobayashi, Koji
Shin-ya, Kazuo
Kitao, Hiroyuki
Wold, Marc S
Branzei, Dana
Neuman, Keir C
Brosh, Robert M
author_sort Bharti, Sanjay Kumar
collection PubMed
description 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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spelling pubmed-61595162018-10-02 A minimal threshold of FANCJ helicase activity is required for its response to replication stress or double-strand break repair Bharti, Sanjay Kumar Sommers, Joshua A Awate, Sanket Bellani, Marina A Khan, Irfan Bradley, Lynda King, Graeme A Seol, Yeonee Vidhyasagar, Venkatasubramanian Wu, Yuliang Abe, Takuye Kobayashi, Koji Shin-ya, Kazuo Kitao, Hiroyuki Wold, Marc S Branzei, Dana Neuman, Keir C Brosh, Robert M Nucleic Acids Res Nucleic Acid Enzymes 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. Oxford University Press 2018-07-06 2018-05-21 /pmc/articles/PMC6159516/ /pubmed/29788478 http://dx.doi.org/10.1093/nar/gky403 Text en Published by Oxford University Press on behalf of Nucleic Acids Research 2018. This work is written by (a) US Government employee(s) and is in the public domain in the US.
spellingShingle Nucleic Acid Enzymes
Bharti, Sanjay Kumar
Sommers, Joshua A
Awate, Sanket
Bellani, Marina A
Khan, Irfan
Bradley, Lynda
King, Graeme A
Seol, Yeonee
Vidhyasagar, Venkatasubramanian
Wu, Yuliang
Abe, Takuye
Kobayashi, Koji
Shin-ya, Kazuo
Kitao, Hiroyuki
Wold, Marc S
Branzei, Dana
Neuman, Keir C
Brosh, Robert M
A minimal threshold of FANCJ helicase activity is required for its response to replication stress or double-strand break repair
title A minimal threshold of FANCJ helicase activity is required for its response to replication stress or double-strand break repair
title_full A minimal threshold of FANCJ helicase activity is required for its response to replication stress or double-strand break repair
title_fullStr A minimal threshold of FANCJ helicase activity is required for its response to replication stress or double-strand break repair
title_full_unstemmed A minimal threshold of FANCJ helicase activity is required for its response to replication stress or double-strand break repair
title_short A minimal threshold of FANCJ helicase activity is required for its response to replication stress or double-strand break repair
title_sort minimal threshold of fancj helicase activity is required for its response to replication stress or double-strand break repair
topic Nucleic Acid Enzymes
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6159516/
https://www.ncbi.nlm.nih.gov/pubmed/29788478
http://dx.doi.org/10.1093/nar/gky403
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