Cargando…
Chemical Incorporation of Chain-Terminating Nucleoside Analogs as 3′-Blocking DNA Damage and Their Removal by Human ERCC1-XPF Endonuclease
Nucleoside/nucleotide analogs that lack the 3′-hydroxy group are widely utilized for HIV therapy. These chain-terminating nucleoside analogs (CTNAs) block DNA synthesis after their incorporation into growing DNA, leading to the antiviral effects. However, they are also considered to be DNA damaging...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6273010/ https://www.ncbi.nlm.nih.gov/pubmed/27294910 http://dx.doi.org/10.3390/molecules21060766 |
_version_ | 1783377285984813056 |
---|---|
author | Yamamoto, Junpei Takahata, Chiaki Kuraoka, Isao Hirota, Kouji Iwai, Shigenori |
author_facet | Yamamoto, Junpei Takahata, Chiaki Kuraoka, Isao Hirota, Kouji Iwai, Shigenori |
author_sort | Yamamoto, Junpei |
collection | PubMed |
description | Nucleoside/nucleotide analogs that lack the 3′-hydroxy group are widely utilized for HIV therapy. These chain-terminating nucleoside analogs (CTNAs) block DNA synthesis after their incorporation into growing DNA, leading to the antiviral effects. However, they are also considered to be DNA damaging agents, and tyrosyl-DNA phosphodiesterase 1, a DNA repair enzyme, is reportedly able to remove such CTNA-modifications of DNA. Here, we have synthesized phosphoramidite building blocks of representative CTNAs, such as acyclovir, abacavir, carbovir, and lamivudine, and oligonucleotides with the 3′-CTNAs were successfully synthesized on solid supports. Using the chemically synthesized oligonucleotides, we investigated the excision of the 3′-CTNAs in DNA by the human excision repair cross complementing protein 1-xeroderma pigmentosum group F (ERCC1-XPF) endonuclease, which is one of the main components of the nucleotide excision repair pathway. A biochemical analysis demonstrated that the ERCC1-XPF endonuclease cleaved 2–7 nt upstream from the 3′-blocking CTNAs, and that DNA synthesis by the Klenow fragment was resumed after the removal of the CTNAs, suggesting that ERCC1-XPF participates in the repair of the CTNA-induced DNA damage. |
format | Online Article Text |
id | pubmed-6273010 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-62730102018-12-28 Chemical Incorporation of Chain-Terminating Nucleoside Analogs as 3′-Blocking DNA Damage and Their Removal by Human ERCC1-XPF Endonuclease Yamamoto, Junpei Takahata, Chiaki Kuraoka, Isao Hirota, Kouji Iwai, Shigenori Molecules Article Nucleoside/nucleotide analogs that lack the 3′-hydroxy group are widely utilized for HIV therapy. These chain-terminating nucleoside analogs (CTNAs) block DNA synthesis after their incorporation into growing DNA, leading to the antiviral effects. However, they are also considered to be DNA damaging agents, and tyrosyl-DNA phosphodiesterase 1, a DNA repair enzyme, is reportedly able to remove such CTNA-modifications of DNA. Here, we have synthesized phosphoramidite building blocks of representative CTNAs, such as acyclovir, abacavir, carbovir, and lamivudine, and oligonucleotides with the 3′-CTNAs were successfully synthesized on solid supports. Using the chemically synthesized oligonucleotides, we investigated the excision of the 3′-CTNAs in DNA by the human excision repair cross complementing protein 1-xeroderma pigmentosum group F (ERCC1-XPF) endonuclease, which is one of the main components of the nucleotide excision repair pathway. A biochemical analysis demonstrated that the ERCC1-XPF endonuclease cleaved 2–7 nt upstream from the 3′-blocking CTNAs, and that DNA synthesis by the Klenow fragment was resumed after the removal of the CTNAs, suggesting that ERCC1-XPF participates in the repair of the CTNA-induced DNA damage. MDPI 2016-06-11 /pmc/articles/PMC6273010/ /pubmed/27294910 http://dx.doi.org/10.3390/molecules21060766 Text en © 2016 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Yamamoto, Junpei Takahata, Chiaki Kuraoka, Isao Hirota, Kouji Iwai, Shigenori Chemical Incorporation of Chain-Terminating Nucleoside Analogs as 3′-Blocking DNA Damage and Their Removal by Human ERCC1-XPF Endonuclease |
title | Chemical Incorporation of Chain-Terminating Nucleoside Analogs as 3′-Blocking DNA Damage and Their Removal by Human ERCC1-XPF Endonuclease |
title_full | Chemical Incorporation of Chain-Terminating Nucleoside Analogs as 3′-Blocking DNA Damage and Their Removal by Human ERCC1-XPF Endonuclease |
title_fullStr | Chemical Incorporation of Chain-Terminating Nucleoside Analogs as 3′-Blocking DNA Damage and Their Removal by Human ERCC1-XPF Endonuclease |
title_full_unstemmed | Chemical Incorporation of Chain-Terminating Nucleoside Analogs as 3′-Blocking DNA Damage and Their Removal by Human ERCC1-XPF Endonuclease |
title_short | Chemical Incorporation of Chain-Terminating Nucleoside Analogs as 3′-Blocking DNA Damage and Their Removal by Human ERCC1-XPF Endonuclease |
title_sort | chemical incorporation of chain-terminating nucleoside analogs as 3′-blocking dna damage and their removal by human ercc1-xpf endonuclease |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6273010/ https://www.ncbi.nlm.nih.gov/pubmed/27294910 http://dx.doi.org/10.3390/molecules21060766 |
work_keys_str_mv | AT yamamotojunpei chemicalincorporationofchainterminatingnucleosideanalogsas3blockingdnadamageandtheirremovalbyhumanercc1xpfendonuclease AT takahatachiaki chemicalincorporationofchainterminatingnucleosideanalogsas3blockingdnadamageandtheirremovalbyhumanercc1xpfendonuclease AT kuraokaisao chemicalincorporationofchainterminatingnucleosideanalogsas3blockingdnadamageandtheirremovalbyhumanercc1xpfendonuclease AT hirotakouji chemicalincorporationofchainterminatingnucleosideanalogsas3blockingdnadamageandtheirremovalbyhumanercc1xpfendonuclease AT iwaishigenori chemicalincorporationofchainterminatingnucleosideanalogsas3blockingdnadamageandtheirremovalbyhumanercc1xpfendonuclease |