Metastasis Suppressor NME1 Modulates Choice of Double-Strand Break Repair Pathways in Melanoma Cells by Enhancing Alternative NHEJ while Inhibiting NHEJ and HR
Reduced NME1 expression in melanoma cell lines, mouse models of melanoma, and melanoma specimens in human patients is associated with increased metastatic activity. Herein, we investigate the role of NME1 in repair of double-stranded breaks (DSBs) and choice of double-strand break repair (DSBR) path...
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doaj-a98c4d2795b5461fac241b68b2eefa072020-11-25T03:46:30ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-08-01215896589610.3390/ijms21165896Metastasis Suppressor NME1 Modulates Choice of Double-Strand Break Repair Pathways in Melanoma Cells by Enhancing Alternative NHEJ while Inhibiting NHEJ and HRGemma Puts0Stuart Jarrett1Mary Leonard2Nicolette Matsangos3Devin Snyder4Ying Wang5Richard Vincent6Benjamin Portney7Rachel Abbotts8Lena McLaughlin9Michal Zalzman10Feyruz Rassool11David Kaetzel12Department of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USADepartment of Toxicology and Cancer Biology, College of Medicine, University of Kentucky, Lexington, KY 40536, USADepartment of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USADepartment of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USADepartment of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USADepartment of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USADepartment of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USADepartment of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USADepartment of Radiation Oncology, School of Medicine, University of Maryland, Baltimore, MD 21201, USADepartment of Radiation Oncology, School of Medicine, University of Maryland, Baltimore, MD 21201, USADepartment of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USADepartment of Radiation Oncology, School of Medicine, University of Maryland, Baltimore, MD 21201, USADepartment of Biochemistry and Molecular Biology, School of Medicine, University of Maryland, Baltimore, MD 21201, USAReduced NME1 expression in melanoma cell lines, mouse models of melanoma, and melanoma specimens in human patients is associated with increased metastatic activity. Herein, we investigate the role of NME1 in repair of double-stranded breaks (DSBs) and choice of double-strand break repair (DSBR) pathways in melanoma cells. Using chromatin immunoprecipitation, NME1 was shown to be recruited rapidly and directly to DSBs generated by the homing endonuclease I-PpoI. NME1 was recruited to DSBs within 30 min, in concert with recruitment of ataxia-telangiectasia mutated (ATM) protein, an early step in DSBR complex formation, as well as loss of histone 2B. NME1 was detected up to 5 kb from the break site after DSB induction, suggesting a role in extending chromatin reorganization away from the repair site. shRNA-mediated silencing of NME1 expression led to increases in the homologous recombination (HR) and non-homologous end-joining (NHEJ) pathways of double-strand break repair (DSBR), and reduction in the low fidelity, alternative-NHEJ (A-NHEJ) pathway. These findings suggest low expression of NME1 drives DSBR towards higher fidelity pathways, conferring enhanced genomic stability necessary for rapid and error-free proliferation in invasive and metastatic cells. The novel mechanism highlighted in the current study appears likely to impact metastatic potential and therapy-resistance in advanced melanoma and other cancers.https://www.mdpi.com/1422-0067/21/16/5896DNA repaircancermelanomametastasisDNA double strand break repairnon-homologous end-joining |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Gemma Puts Stuart Jarrett Mary Leonard Nicolette Matsangos Devin Snyder Ying Wang Richard Vincent Benjamin Portney Rachel Abbotts Lena McLaughlin Michal Zalzman Feyruz Rassool David Kaetzel |
spellingShingle |
Gemma Puts Stuart Jarrett Mary Leonard Nicolette Matsangos Devin Snyder Ying Wang Richard Vincent Benjamin Portney Rachel Abbotts Lena McLaughlin Michal Zalzman Feyruz Rassool David Kaetzel Metastasis Suppressor NME1 Modulates Choice of Double-Strand Break Repair Pathways in Melanoma Cells by Enhancing Alternative NHEJ while Inhibiting NHEJ and HR International Journal of Molecular Sciences DNA repair cancer melanoma metastasis DNA double strand break repair non-homologous end-joining |
author_facet |
Gemma Puts Stuart Jarrett Mary Leonard Nicolette Matsangos Devin Snyder Ying Wang Richard Vincent Benjamin Portney Rachel Abbotts Lena McLaughlin Michal Zalzman Feyruz Rassool David Kaetzel |
author_sort |
Gemma Puts |
title |
Metastasis Suppressor NME1 Modulates Choice of Double-Strand Break Repair Pathways in Melanoma Cells by Enhancing Alternative NHEJ while Inhibiting NHEJ and HR |
title_short |
Metastasis Suppressor NME1 Modulates Choice of Double-Strand Break Repair Pathways in Melanoma Cells by Enhancing Alternative NHEJ while Inhibiting NHEJ and HR |
title_full |
Metastasis Suppressor NME1 Modulates Choice of Double-Strand Break Repair Pathways in Melanoma Cells by Enhancing Alternative NHEJ while Inhibiting NHEJ and HR |
title_fullStr |
Metastasis Suppressor NME1 Modulates Choice of Double-Strand Break Repair Pathways in Melanoma Cells by Enhancing Alternative NHEJ while Inhibiting NHEJ and HR |
title_full_unstemmed |
Metastasis Suppressor NME1 Modulates Choice of Double-Strand Break Repair Pathways in Melanoma Cells by Enhancing Alternative NHEJ while Inhibiting NHEJ and HR |
title_sort |
metastasis suppressor nme1 modulates choice of double-strand break repair pathways in melanoma cells by enhancing alternative nhej while inhibiting nhej and hr |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2020-08-01 |
description |
Reduced NME1 expression in melanoma cell lines, mouse models of melanoma, and melanoma specimens in human patients is associated with increased metastatic activity. Herein, we investigate the role of NME1 in repair of double-stranded breaks (DSBs) and choice of double-strand break repair (DSBR) pathways in melanoma cells. Using chromatin immunoprecipitation, NME1 was shown to be recruited rapidly and directly to DSBs generated by the homing endonuclease I-PpoI. NME1 was recruited to DSBs within 30 min, in concert with recruitment of ataxia-telangiectasia mutated (ATM) protein, an early step in DSBR complex formation, as well as loss of histone 2B. NME1 was detected up to 5 kb from the break site after DSB induction, suggesting a role in extending chromatin reorganization away from the repair site. shRNA-mediated silencing of NME1 expression led to increases in the homologous recombination (HR) and non-homologous end-joining (NHEJ) pathways of double-strand break repair (DSBR), and reduction in the low fidelity, alternative-NHEJ (A-NHEJ) pathway. These findings suggest low expression of NME1 drives DSBR towards higher fidelity pathways, conferring enhanced genomic stability necessary for rapid and error-free proliferation in invasive and metastatic cells. The novel mechanism highlighted in the current study appears likely to impact metastatic potential and therapy-resistance in advanced melanoma and other cancers. |
topic |
DNA repair cancer melanoma metastasis DNA double strand break repair non-homologous end-joining |
url |
https://www.mdpi.com/1422-0067/21/16/5896 |
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