A clickable psoralen to directly quantify DNA interstrand crosslinking and repair

[Display omitted] DNA interstrand crosslinks (ICLs) represent physical obstacles to advancing replication forks and transcription complexes. A range of ICL-inducing agents have successfully been incorporated into cancer therapeutics. While studies have adopted UVA-activated psoralens as model ICL-in...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Bioorganic & medicinal chemistry 2016-03, Vol.24 (5), p.1071-1078
Hauptverfasser: Evison, Benjamin J., Actis, Marcelo L., Fujii, Naoaki
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 1078
container_issue 5
container_start_page 1071
container_title Bioorganic & medicinal chemistry
container_volume 24
creator Evison, Benjamin J.
Actis, Marcelo L.
Fujii, Naoaki
description [Display omitted] DNA interstrand crosslinks (ICLs) represent physical obstacles to advancing replication forks and transcription complexes. A range of ICL-inducing agents have successfully been incorporated into cancer therapeutics. While studies have adopted UVA-activated psoralens as model ICL-inducing agents for investigating ICL repair, direct detection of the lesion has often been tempered by tagging the psoralen scaffold with a relatively large reporter group that may perturb the biological activity of the parent psoralen. Here a minimally-modified psoralen probe was prepared featuring a small alkyne handle suitable for click chemistry. The psoralen probe, designated 8-propargyloxypsoralen (8-POP), can be activated by UVA in vitro to generate ICLs that are susceptible to post-labeling with an azide-tagged fluorescent reporter via a copper-catalyzed reaction. A modified alkaline comet assay demonstrated that UVA-activated 8-POP proficiently generated ICLs in cells. Cellular 8-POP–DNA lesions were amenable to click-mediated ligation to fluorescent reporters in situ, which permitted their detection and quantitation by fluorescence microscopy and flow cytometry. Small molecule DNA repair inhibitors to 8-POP-treated cells attenuated the removal of 8-POP–DNA lesions, validating 8-POP as an appropriate probe for investigating cellular ICL repair. The post-labeling strategy applied in this study is inexpensive, rapid and highly modular in nature with the potential for multiple applications in DNA repair studies.
doi_str_mv 10.1016/j.bmc.2016.01.032
format Article
fullrecord <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4762217</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0968089616300311</els_id><sourcerecordid>1790942114</sourcerecordid><originalsourceid>FETCH-LOGICAL-c550t-e14681b2276749caeee1676886c7a834595d33160d74d242fe86c759121ee5ca3</originalsourceid><addsrcrecordid>eNqNkU2LFDEQhoMo7rj6A7xIjl66TeWrEwRhWD9hUQQ9h0y6Zs1sJj2b9CzMvzfjrItexFOFqrde3tRDyHNgPTDQrzb9aht63p49g54J_oAsQGrZCWHhIVkwq03HjNVn5EmtG8YYlxYekzOujRBcygX5uqQhxXDtVwnprk7FJ8x0nugYC4Y5HejN3uc5rg_07ecljXnGUufi80hDmWpNMV_HfEWPjYI7H8tT8mjtU8Vnd_WcfH__7tvFx-7yy4dPF8vLLijF5g5bUAMrzgc9SBs8IoIetDE6DN4IqawahQDNxkGOXPI1HifKAgdEFbw4J29Ovrv9aotjwNxiJbcrcevLwU0-ur8nOf5wV9Otk4PmHIZm8PLOoEw3e6yz28YaMCWfcdpXB4NlVnIA-R9SrZRhhqkmhZP013kKru8TAXNHam7jGjV3pOYYuEat7bz48yv3G78xNcHrkwDbQW8jFldDxBzwRMmNU_yH_U9ez6fz</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1765580805</pqid></control><display><type>article</type><title>A clickable psoralen to directly quantify DNA interstrand crosslinking and repair</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>Evison, Benjamin J. ; Actis, Marcelo L. ; Fujii, Naoaki</creator><creatorcontrib>Evison, Benjamin J. ; Actis, Marcelo L. ; Fujii, Naoaki</creatorcontrib><description>[Display omitted] DNA interstrand crosslinks (ICLs) represent physical obstacles to advancing replication forks and transcription complexes. A range of ICL-inducing agents have successfully been incorporated into cancer therapeutics. While studies have adopted UVA-activated psoralens as model ICL-inducing agents for investigating ICL repair, direct detection of the lesion has often been tempered by tagging the psoralen scaffold with a relatively large reporter group that may perturb the biological activity of the parent psoralen. Here a minimally-modified psoralen probe was prepared featuring a small alkyne handle suitable for click chemistry. The psoralen probe, designated 8-propargyloxypsoralen (8-POP), can be activated by UVA in vitro to generate ICLs that are susceptible to post-labeling with an azide-tagged fluorescent reporter via a copper-catalyzed reaction. A modified alkaline comet assay demonstrated that UVA-activated 8-POP proficiently generated ICLs in cells. Cellular 8-POP–DNA lesions were amenable to click-mediated ligation to fluorescent reporters in situ, which permitted their detection and quantitation by fluorescence microscopy and flow cytometry. Small molecule DNA repair inhibitors to 8-POP-treated cells attenuated the removal of 8-POP–DNA lesions, validating 8-POP as an appropriate probe for investigating cellular ICL repair. The post-labeling strategy applied in this study is inexpensive, rapid and highly modular in nature with the potential for multiple applications in DNA repair studies.</description><identifier>ISSN: 0968-0896</identifier><identifier>EISSN: 1464-3391</identifier><identifier>DOI: 10.1016/j.bmc.2016.01.032</identifier><identifier>PMID: 26833244</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>Alkynes - chemistry ; Azides - chemistry ; Chemical screening ; Click Chemistry ; Copper - chemistry ; DNA Adducts - analysis ; DNA Repair ; Ficusin - chemistry ; Fluorescent Dyes - chemistry ; HeLa Cells ; Humans ; Imaging ; Interstrand DNA crosslink ; Psoralen ; Ultraviolet Rays</subject><ispartof>Bioorganic &amp; medicinal chemistry, 2016-03, Vol.24 (5), p.1071-1078</ispartof><rights>2016 Elsevier Ltd</rights><rights>Copyright © 2016 Elsevier Ltd. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c550t-e14681b2276749caeee1676886c7a834595d33160d74d242fe86c759121ee5ca3</citedby><cites>FETCH-LOGICAL-c550t-e14681b2276749caeee1676886c7a834595d33160d74d242fe86c759121ee5ca3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.bmc.2016.01.032$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,780,784,885,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26833244$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Evison, Benjamin J.</creatorcontrib><creatorcontrib>Actis, Marcelo L.</creatorcontrib><creatorcontrib>Fujii, Naoaki</creatorcontrib><title>A clickable psoralen to directly quantify DNA interstrand crosslinking and repair</title><title>Bioorganic &amp; medicinal chemistry</title><addtitle>Bioorg Med Chem</addtitle><description>[Display omitted] DNA interstrand crosslinks (ICLs) represent physical obstacles to advancing replication forks and transcription complexes. A range of ICL-inducing agents have successfully been incorporated into cancer therapeutics. While studies have adopted UVA-activated psoralens as model ICL-inducing agents for investigating ICL repair, direct detection of the lesion has often been tempered by tagging the psoralen scaffold with a relatively large reporter group that may perturb the biological activity of the parent psoralen. Here a minimally-modified psoralen probe was prepared featuring a small alkyne handle suitable for click chemistry. The psoralen probe, designated 8-propargyloxypsoralen (8-POP), can be activated by UVA in vitro to generate ICLs that are susceptible to post-labeling with an azide-tagged fluorescent reporter via a copper-catalyzed reaction. A modified alkaline comet assay demonstrated that UVA-activated 8-POP proficiently generated ICLs in cells. Cellular 8-POP–DNA lesions were amenable to click-mediated ligation to fluorescent reporters in situ, which permitted their detection and quantitation by fluorescence microscopy and flow cytometry. Small molecule DNA repair inhibitors to 8-POP-treated cells attenuated the removal of 8-POP–DNA lesions, validating 8-POP as an appropriate probe for investigating cellular ICL repair. The post-labeling strategy applied in this study is inexpensive, rapid and highly modular in nature with the potential for multiple applications in DNA repair studies.</description><subject>Alkynes - chemistry</subject><subject>Azides - chemistry</subject><subject>Chemical screening</subject><subject>Click Chemistry</subject><subject>Copper - chemistry</subject><subject>DNA Adducts - analysis</subject><subject>DNA Repair</subject><subject>Ficusin - chemistry</subject><subject>Fluorescent Dyes - chemistry</subject><subject>HeLa Cells</subject><subject>Humans</subject><subject>Imaging</subject><subject>Interstrand DNA crosslink</subject><subject>Psoralen</subject><subject>Ultraviolet Rays</subject><issn>0968-0896</issn><issn>1464-3391</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkU2LFDEQhoMo7rj6A7xIjl66TeWrEwRhWD9hUQQ9h0y6Zs1sJj2b9CzMvzfjrItexFOFqrde3tRDyHNgPTDQrzb9aht63p49g54J_oAsQGrZCWHhIVkwq03HjNVn5EmtG8YYlxYekzOujRBcygX5uqQhxXDtVwnprk7FJ8x0nugYC4Y5HejN3uc5rg_07ecljXnGUufi80hDmWpNMV_HfEWPjYI7H8tT8mjtU8Vnd_WcfH__7tvFx-7yy4dPF8vLLijF5g5bUAMrzgc9SBs8IoIetDE6DN4IqawahQDNxkGOXPI1HifKAgdEFbw4J29Ovrv9aotjwNxiJbcrcevLwU0-ur8nOf5wV9Otk4PmHIZm8PLOoEw3e6yz28YaMCWfcdpXB4NlVnIA-R9SrZRhhqkmhZP013kKru8TAXNHam7jGjV3pOYYuEat7bz48yv3G78xNcHrkwDbQW8jFldDxBzwRMmNU_yH_U9ez6fz</recordid><startdate>20160301</startdate><enddate>20160301</enddate><creator>Evison, Benjamin J.</creator><creator>Actis, Marcelo L.</creator><creator>Fujii, Naoaki</creator><general>Elsevier Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7QO</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>5PM</scope></search><sort><creationdate>20160301</creationdate><title>A clickable psoralen to directly quantify DNA interstrand crosslinking and repair</title><author>Evison, Benjamin J. ; Actis, Marcelo L. ; Fujii, Naoaki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c550t-e14681b2276749caeee1676886c7a834595d33160d74d242fe86c759121ee5ca3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Alkynes - chemistry</topic><topic>Azides - chemistry</topic><topic>Chemical screening</topic><topic>Click Chemistry</topic><topic>Copper - chemistry</topic><topic>DNA Adducts - analysis</topic><topic>DNA Repair</topic><topic>Ficusin - chemistry</topic><topic>Fluorescent Dyes - chemistry</topic><topic>HeLa Cells</topic><topic>Humans</topic><topic>Imaging</topic><topic>Interstrand DNA crosslink</topic><topic>Psoralen</topic><topic>Ultraviolet Rays</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Evison, Benjamin J.</creatorcontrib><creatorcontrib>Actis, Marcelo L.</creatorcontrib><creatorcontrib>Fujii, Naoaki</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Biotechnology Research Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Bioorganic &amp; medicinal chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Evison, Benjamin J.</au><au>Actis, Marcelo L.</au><au>Fujii, Naoaki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A clickable psoralen to directly quantify DNA interstrand crosslinking and repair</atitle><jtitle>Bioorganic &amp; medicinal chemistry</jtitle><addtitle>Bioorg Med Chem</addtitle><date>2016-03-01</date><risdate>2016</risdate><volume>24</volume><issue>5</issue><spage>1071</spage><epage>1078</epage><pages>1071-1078</pages><issn>0968-0896</issn><eissn>1464-3391</eissn><abstract>[Display omitted] DNA interstrand crosslinks (ICLs) represent physical obstacles to advancing replication forks and transcription complexes. A range of ICL-inducing agents have successfully been incorporated into cancer therapeutics. While studies have adopted UVA-activated psoralens as model ICL-inducing agents for investigating ICL repair, direct detection of the lesion has often been tempered by tagging the psoralen scaffold with a relatively large reporter group that may perturb the biological activity of the parent psoralen. Here a minimally-modified psoralen probe was prepared featuring a small alkyne handle suitable for click chemistry. The psoralen probe, designated 8-propargyloxypsoralen (8-POP), can be activated by UVA in vitro to generate ICLs that are susceptible to post-labeling with an azide-tagged fluorescent reporter via a copper-catalyzed reaction. A modified alkaline comet assay demonstrated that UVA-activated 8-POP proficiently generated ICLs in cells. Cellular 8-POP–DNA lesions were amenable to click-mediated ligation to fluorescent reporters in situ, which permitted their detection and quantitation by fluorescence microscopy and flow cytometry. Small molecule DNA repair inhibitors to 8-POP-treated cells attenuated the removal of 8-POP–DNA lesions, validating 8-POP as an appropriate probe for investigating cellular ICL repair. The post-labeling strategy applied in this study is inexpensive, rapid and highly modular in nature with the potential for multiple applications in DNA repair studies.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>26833244</pmid><doi>10.1016/j.bmc.2016.01.032</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 0968-0896
ispartof Bioorganic & medicinal chemistry, 2016-03, Vol.24 (5), p.1071-1078
issn 0968-0896
1464-3391
language eng
recordid cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4762217
source MEDLINE; Access via ScienceDirect (Elsevier)
subjects Alkynes - chemistry
Azides - chemistry
Chemical screening
Click Chemistry
Copper - chemistry
DNA Adducts - analysis
DNA Repair
Ficusin - chemistry
Fluorescent Dyes - chemistry
HeLa Cells
Humans
Imaging
Interstrand DNA crosslink
Psoralen
Ultraviolet Rays
title A clickable psoralen to directly quantify DNA interstrand crosslinking and repair
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-24T18%3A23%3A58IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20clickable%20psoralen%20to%20directly%20quantify%20DNA%20interstrand%20crosslinking%20and%20repair&rft.jtitle=Bioorganic%20&%20medicinal%20chemistry&rft.au=Evison,%20Benjamin%20J.&rft.date=2016-03-01&rft.volume=24&rft.issue=5&rft.spage=1071&rft.epage=1078&rft.pages=1071-1078&rft.issn=0968-0896&rft.eissn=1464-3391&rft_id=info:doi/10.1016/j.bmc.2016.01.032&rft_dat=%3Cproquest_pubme%3E1790942114%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1765580805&rft_id=info:pmid/26833244&rft_els_id=S0968089616300311&rfr_iscdi=true