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Phage N15 protelomerase resolves its tos recognition site into hairpin telomeres within mammalian cells

Phage N15 protelomerase (TelN) cleaves double-stranded circular DNA containing a telomerase-occupancy-site (tos) and rejoins the resulting linear-ends to form closed-hairpin-telomeres in Escherichia coli (E. coli). ContinuedTelN expression is essential to support resolution of the linear structure....

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Main Authors: Liew, Pei Sheng, Chen, Qingwen, Ng, Allan Wee Ren, Chew, Yee Choy, Ravin, Nikolai V., Sim, Edmund Ui Hang, Lee, Choon-Weng, Narayanan, Kumaran
Format: E-Article
Language:English
Published: Elsevier Ltd. 2019
Subjects:
Q Science (General)
QH426 Genetics
Online Access:http://ir.unimas.my/id/eprint/25890/1/Phage%20N15%20protelomerase%20resolves%20its%20tos%20recognition%20site%20into%20hairpin%20-%20Copy.pdf
http://ir.unimas.my/id/eprint/25890/
https://www.sciencedirect.com/science/article/pii/S0003269719302209
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spelling my.unimas.ir.258902020-06-23T03:44:04Z http://ir.unimas.my/id/eprint/25890/ Phage N15 protelomerase resolves its tos recognition site into hairpin telomeres within mammalian cells Liew, Pei Sheng Chen, Qingwen Ng, Allan Wee Ren Chew, Yee Choy Ravin, Nikolai V. Sim, Edmund Ui Hang Lee, Choon-Weng Narayanan, Kumaran Q Science (General) QH426 Genetics Phage N15 protelomerase (TelN) cleaves double-stranded circular DNA containing a telomerase-occupancy-site (tos) and rejoins the resulting linear-ends to form closed-hairpin-telomeres in Escherichia coli (E. coli). ContinuedTelN expression is essential to support resolution of the linear structure. In mammalian cells, no enzyme withTelN-like activities has been found. In this work, we show that phage TelN, expressed transiently and stably in human and mouse cells, recapitulates its native activities in these exogenous environments. We found TelN to accurately resolve tos-DNA in vitro and in vivo within human and mouse cells into linear DNA-containing terminal telomeres that are resistant to RecBCD degradation, a hallmark of protelomerase processing. In stable cells, TelN activity was detectable for at least 60 days, which suggests the possibility of limited silencing of its expression.Correspondingly, linear plasmid containing a 100 kb human β-globin gene expressed for at least 120 h in non-β-globin-expressing mouse cells with TelN presence. Our results demonstrate TelN is able to cut and heal DNA ashairpin-telomeres within mammalian cells, providing a tool for creating novel structures by DNA resolution in these hosts. The TelN protelomerase may be useful for exploring novel technologies for genome interrogationand chromosome engineering. Elsevier Ltd. 2019-07-12 E-Article PeerReviewed text en http://ir.unimas.my/id/eprint/25890/1/Phage%20N15%20protelomerase%20resolves%20its%20tos%20recognition%20site%20into%20hairpin%20-%20Copy.pdf Liew, Pei Sheng and Chen, Qingwen and Ng, Allan Wee Ren and Chew, Yee Choy and Ravin, Nikolai V. and Sim, Edmund Ui Hang and Lee, Choon-Weng and Narayanan, Kumaran (2019) Phage N15 protelomerase resolves its tos recognition site into hairpin telomeres within mammalian cells. Analytical Biochemistry, 583 (113361). pp. 1-10. ISSN 0003-2697 https://www.sciencedirect.com/science/article/pii/S0003269719302209 DOI:org/10.1016/j.ab.2019.113361
institution Universiti Malaysia Sarawak
building Centre for Academic Information Services (CAIS)
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Sarawak
content_source UNIMAS Institutional Repository
url_provider http://ir.unimas.my/
language English
topic Q Science (General)
QH426 Genetics
spellingShingle Q Science (General)
QH426 Genetics
Liew, Pei Sheng
Chen, Qingwen
Ng, Allan Wee Ren
Chew, Yee Choy
Ravin, Nikolai V.
Sim, Edmund Ui Hang
Lee, Choon-Weng
Narayanan, Kumaran
Phage N15 protelomerase resolves its tos recognition site into hairpin telomeres within mammalian cells
description Phage N15 protelomerase (TelN) cleaves double-stranded circular DNA containing a telomerase-occupancy-site (tos) and rejoins the resulting linear-ends to form closed-hairpin-telomeres in Escherichia coli (E. coli). ContinuedTelN expression is essential to support resolution of the linear structure. In mammalian cells, no enzyme withTelN-like activities has been found. In this work, we show that phage TelN, expressed transiently and stably in human and mouse cells, recapitulates its native activities in these exogenous environments. We found TelN to accurately resolve tos-DNA in vitro and in vivo within human and mouse cells into linear DNA-containing terminal telomeres that are resistant to RecBCD degradation, a hallmark of protelomerase processing. In stable cells, TelN activity was detectable for at least 60 days, which suggests the possibility of limited silencing of its expression.Correspondingly, linear plasmid containing a 100 kb human β-globin gene expressed for at least 120 h in non-β-globin-expressing mouse cells with TelN presence. Our results demonstrate TelN is able to cut and heal DNA ashairpin-telomeres within mammalian cells, providing a tool for creating novel structures by DNA resolution in these hosts. The TelN protelomerase may be useful for exploring novel technologies for genome interrogationand chromosome engineering.
format E-Article
author Liew, Pei Sheng
Chen, Qingwen
Ng, Allan Wee Ren
Chew, Yee Choy
Ravin, Nikolai V.
Sim, Edmund Ui Hang
Lee, Choon-Weng
Narayanan, Kumaran
author_facet Liew, Pei Sheng
Chen, Qingwen
Ng, Allan Wee Ren
Chew, Yee Choy
Ravin, Nikolai V.
Sim, Edmund Ui Hang
Lee, Choon-Weng
Narayanan, Kumaran
author_sort Liew, Pei Sheng
title Phage N15 protelomerase resolves its tos recognition site into hairpin telomeres within mammalian cells
title_short Phage N15 protelomerase resolves its tos recognition site into hairpin telomeres within mammalian cells
title_full Phage N15 protelomerase resolves its tos recognition site into hairpin telomeres within mammalian cells
title_fullStr Phage N15 protelomerase resolves its tos recognition site into hairpin telomeres within mammalian cells
title_full_unstemmed Phage N15 protelomerase resolves its tos recognition site into hairpin telomeres within mammalian cells
title_sort phage n15 protelomerase resolves its tos recognition site into hairpin telomeres within mammalian cells
publisher Elsevier Ltd.
publishDate 2019
url http://ir.unimas.my/id/eprint/25890/1/Phage%20N15%20protelomerase%20resolves%20its%20tos%20recognition%20site%20into%20hairpin%20-%20Copy.pdf
http://ir.unimas.my/id/eprint/25890/
https://www.sciencedirect.com/science/article/pii/S0003269719302209
_version_ 1671343271601766400
score 13.211869

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