Escherichia coli β-clamp slows down DNA polymerase I dependent nick translation while accelerating ligation
The nick translation property of DNA polymerase I (Pol I) ensures the maturation of Okazaki fragments by removing primer RNAs and facilitating ligation. However, prolonged nick translation traversing downstream DNA is an energy wasting futile process, as Pol I simultaneously polymerizes and depolyme...
Gespeichert in:
| Veröffentlicht in: | PloS one 2018-06, Vol.13 (6), p.e0199559-e0199559 |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | e0199559 |
|---|---|
| container_issue | 6 |
| container_start_page | e0199559 |
| container_title | PloS one |
| container_volume | 13 |
| creator | Bhardwaj, Amit Ghose, Debarghya Thakur, Krishan Gopal Dutta, Dipak |
| description | The nick translation property of DNA polymerase I (Pol I) ensures the maturation of Okazaki fragments by removing primer RNAs and facilitating ligation. However, prolonged nick translation traversing downstream DNA is an energy wasting futile process, as Pol I simultaneously polymerizes and depolymerizes at the nick sites utilizing energy-rich dNTPs. Using an in vitro assay system, we demonstrate that the β-clamp of the Escherichia coli replisome strongly inhibits nick translation on the DNA substrate. To do so, β-clamp inhibits the strand displacement activity of Pol I by interfering with the interaction between the finger subdomain of Pol I and the downstream primer-template junction. Conversely, β-clamp stimulates the 5' exonuclease property of Pol I to cleave single nucleotides or shorter oligonucleotide flaps. This single nucleotide flap removal at high frequency increases the probability of ligation between the upstream and downstream DNA strands at an early phase, terminating nick translation. Besides β-clamp-mediated ligation helps DNA ligase to seal the nick promptly during the maturation of Okazaki fragments. |
| doi_str_mv | 10.1371/journal.pone.0199559 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_plos_</sourceid><recordid>TN_cdi_plos_journals_2062793146</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><doaj_id>oai_doaj_org_article_240414fb3a7d4bbca4fab5f22815f941</doaj_id><sourcerecordid>2057865460</sourcerecordid><originalsourceid>FETCH-LOGICAL-c526t-d48ec9ee14a9228159358e532368b43cc0808eabfe872203a0fb14fcb2fe636a3</originalsourceid><addsrcrecordid>eNptUktuFDEUbCEQCYEbILDEhs0M_nd7gxSFACNFsIG19dr9esbBYzd2D6Nci4NwJno-iRLEypZfVflVqarqJaNzJmr27jptcoQwH1LEOWXGKGUeVafMCD7TnIrH9-4n1bNSrilVotH6aXXCjeGykea0CpfFrTB7t_JAXAqe_Pk9cwHWAykhbQvp0jaSD1_OyZDCzRozFCQL0uGAscM4kujdDzJmiCXA6FMk25UPSMA5DBN69HFJgl_uZ8-rJz2Egi-O51n1_ePlt4vPs6uvnxYX51czp7geZ51s0BlEJsFw3jBlhGpQCS5000rhHG1og9D22NR8cge0b5nsXct71EKDOKteH3SHkIo9BlUsp5rXRjCpJ8TigOgSXNsh-zXkG5vA2_1DyksLefQuoOWSykm9FVB3sm0dyB5a1e8X641kk9b742-bdo2dm1LJEB6IPpxEv7LL9Mtqyiiv1STw9iiQ088NltGufZniCxAxbXZ7q7rRSmo6Qd_8A_2_O3lAuZxKydjfLcOo3ZXnlmV35bHH8ky0V_eN3JFu2yL-AhOkxNM</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2062793146</pqid></control><display><type>article</type><title>Escherichia coli β-clamp slows down DNA polymerase I dependent nick translation while accelerating ligation</title><source>DOAJ Directory of Open Access Journals</source><source>Public Library of Science (PLoS)</source><source>EZB-FREE-00999 freely available EZB journals</source><source>PubMed Central</source><source>Free Full-Text Journals in Chemistry</source><creator>Bhardwaj, Amit ; Ghose, Debarghya ; Thakur, Krishan Gopal ; Dutta, Dipak</creator><contributor>Maga, Giovanni</contributor><creatorcontrib>Bhardwaj, Amit ; Ghose, Debarghya ; Thakur, Krishan Gopal ; Dutta, Dipak ; Maga, Giovanni</creatorcontrib><description>The nick translation property of DNA polymerase I (Pol I) ensures the maturation of Okazaki fragments by removing primer RNAs and facilitating ligation. However, prolonged nick translation traversing downstream DNA is an energy wasting futile process, as Pol I simultaneously polymerizes and depolymerizes at the nick sites utilizing energy-rich dNTPs. Using an in vitro assay system, we demonstrate that the β-clamp of the Escherichia coli replisome strongly inhibits nick translation on the DNA substrate. To do so, β-clamp inhibits the strand displacement activity of Pol I by interfering with the interaction between the finger subdomain of Pol I and the downstream primer-template junction. Conversely, β-clamp stimulates the 5' exonuclease property of Pol I to cleave single nucleotides or shorter oligonucleotide flaps. This single nucleotide flap removal at high frequency increases the probability of ligation between the upstream and downstream DNA strands at an early phase, terminating nick translation. Besides β-clamp-mediated ligation helps DNA ligase to seal the nick promptly during the maturation of Okazaki fragments.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0199559</identifier><identifier>PMID: 29924849</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Bacillus subtilis ; Bacteria ; Biology and Life Sciences ; Deoxyribonucleic acid ; Depolymerization ; Displacement activity ; DNA ; DNA polymerase ; DNA repair ; DNA-directed DNA polymerase ; E coli ; Escherichia coli ; Exonuclease ; Flaps ; Fragments ; Maturation ; Nucleotides ; Okazaki fragments ; Oligonucleotides ; Physical Sciences ; Polypeptides ; Research and Analysis Methods ; Substrate inhibition ; Translation</subject><ispartof>PloS one, 2018-06, Vol.13 (6), p.e0199559-e0199559</ispartof><rights>2018 Bhardwaj et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2018 Bhardwaj et al 2018 Bhardwaj et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c526t-d48ec9ee14a9228159358e532368b43cc0808eabfe872203a0fb14fcb2fe636a3</citedby><cites>FETCH-LOGICAL-c526t-d48ec9ee14a9228159358e532368b43cc0808eabfe872203a0fb14fcb2fe636a3</cites><orcidid>0000-0002-0458-4109</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6010275/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6010275/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2100,2926,23865,27923,27924,53790,53792,79371,79372</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29924849$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Maga, Giovanni</contributor><creatorcontrib>Bhardwaj, Amit</creatorcontrib><creatorcontrib>Ghose, Debarghya</creatorcontrib><creatorcontrib>Thakur, Krishan Gopal</creatorcontrib><creatorcontrib>Dutta, Dipak</creatorcontrib><title>Escherichia coli β-clamp slows down DNA polymerase I dependent nick translation while accelerating ligation</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>The nick translation property of DNA polymerase I (Pol I) ensures the maturation of Okazaki fragments by removing primer RNAs and facilitating ligation. However, prolonged nick translation traversing downstream DNA is an energy wasting futile process, as Pol I simultaneously polymerizes and depolymerizes at the nick sites utilizing energy-rich dNTPs. Using an in vitro assay system, we demonstrate that the β-clamp of the Escherichia coli replisome strongly inhibits nick translation on the DNA substrate. To do so, β-clamp inhibits the strand displacement activity of Pol I by interfering with the interaction between the finger subdomain of Pol I and the downstream primer-template junction. Conversely, β-clamp stimulates the 5' exonuclease property of Pol I to cleave single nucleotides or shorter oligonucleotide flaps. This single nucleotide flap removal at high frequency increases the probability of ligation between the upstream and downstream DNA strands at an early phase, terminating nick translation. Besides β-clamp-mediated ligation helps DNA ligase to seal the nick promptly during the maturation of Okazaki fragments.</description><subject>Bacillus subtilis</subject><subject>Bacteria</subject><subject>Biology and Life Sciences</subject><subject>Deoxyribonucleic acid</subject><subject>Depolymerization</subject><subject>Displacement activity</subject><subject>DNA</subject><subject>DNA polymerase</subject><subject>DNA repair</subject><subject>DNA-directed DNA polymerase</subject><subject>E coli</subject><subject>Escherichia coli</subject><subject>Exonuclease</subject><subject>Flaps</subject><subject>Fragments</subject><subject>Maturation</subject><subject>Nucleotides</subject><subject>Okazaki fragments</subject><subject>Oligonucleotides</subject><subject>Physical Sciences</subject><subject>Polypeptides</subject><subject>Research and Analysis Methods</subject><subject>Substrate inhibition</subject><subject>Translation</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNptUktuFDEUbCEQCYEbILDEhs0M_nd7gxSFACNFsIG19dr9esbBYzd2D6Nci4NwJno-iRLEypZfVflVqarqJaNzJmr27jptcoQwH1LEOWXGKGUeVafMCD7TnIrH9-4n1bNSrilVotH6aXXCjeGykea0CpfFrTB7t_JAXAqe_Pk9cwHWAykhbQvp0jaSD1_OyZDCzRozFCQL0uGAscM4kujdDzJmiCXA6FMk25UPSMA5DBN69HFJgl_uZ8-rJz2Egi-O51n1_ePlt4vPs6uvnxYX51czp7geZ51s0BlEJsFw3jBlhGpQCS5000rhHG1og9D22NR8cge0b5nsXct71EKDOKteH3SHkIo9BlUsp5rXRjCpJ8TigOgSXNsh-zXkG5vA2_1DyksLefQuoOWSykm9FVB3sm0dyB5a1e8X641kk9b742-bdo2dm1LJEB6IPpxEv7LL9Mtqyiiv1STw9iiQ088NltGufZniCxAxbXZ7q7rRSmo6Qd_8A_2_O3lAuZxKydjfLcOo3ZXnlmV35bHH8ky0V_eN3JFu2yL-AhOkxNM</recordid><startdate>20180620</startdate><enddate>20180620</enddate><creator>Bhardwaj, Amit</creator><creator>Ghose, Debarghya</creator><creator>Thakur, Krishan Gopal</creator><creator>Dutta, Dipak</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0002-0458-4109</orcidid></search><sort><creationdate>20180620</creationdate><title>Escherichia coli β-clamp slows down DNA polymerase I dependent nick translation while accelerating ligation</title><author>Bhardwaj, Amit ; Ghose, Debarghya ; Thakur, Krishan Gopal ; Dutta, Dipak</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c526t-d48ec9ee14a9228159358e532368b43cc0808eabfe872203a0fb14fcb2fe636a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Bacillus subtilis</topic><topic>Bacteria</topic><topic>Biology and Life Sciences</topic><topic>Deoxyribonucleic acid</topic><topic>Depolymerization</topic><topic>Displacement activity</topic><topic>DNA</topic><topic>DNA polymerase</topic><topic>DNA repair</topic><topic>DNA-directed DNA polymerase</topic><topic>E coli</topic><topic>Escherichia coli</topic><topic>Exonuclease</topic><topic>Flaps</topic><topic>Fragments</topic><topic>Maturation</topic><topic>Nucleotides</topic><topic>Okazaki fragments</topic><topic>Oligonucleotides</topic><topic>Physical Sciences</topic><topic>Polypeptides</topic><topic>Research and Analysis Methods</topic><topic>Substrate inhibition</topic><topic>Translation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bhardwaj, Amit</creatorcontrib><creatorcontrib>Ghose, Debarghya</creatorcontrib><creatorcontrib>Thakur, Krishan Gopal</creatorcontrib><creatorcontrib>Dutta, Dipak</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - Academic</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Materials Science Collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bhardwaj, Amit</au><au>Ghose, Debarghya</au><au>Thakur, Krishan Gopal</au><au>Dutta, Dipak</au><au>Maga, Giovanni</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Escherichia coli β-clamp slows down DNA polymerase I dependent nick translation while accelerating ligation</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2018-06-20</date><risdate>2018</risdate><volume>13</volume><issue>6</issue><spage>e0199559</spage><epage>e0199559</epage><pages>e0199559-e0199559</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>The nick translation property of DNA polymerase I (Pol I) ensures the maturation of Okazaki fragments by removing primer RNAs and facilitating ligation. However, prolonged nick translation traversing downstream DNA is an energy wasting futile process, as Pol I simultaneously polymerizes and depolymerizes at the nick sites utilizing energy-rich dNTPs. Using an in vitro assay system, we demonstrate that the β-clamp of the Escherichia coli replisome strongly inhibits nick translation on the DNA substrate. To do so, β-clamp inhibits the strand displacement activity of Pol I by interfering with the interaction between the finger subdomain of Pol I and the downstream primer-template junction. Conversely, β-clamp stimulates the 5' exonuclease property of Pol I to cleave single nucleotides or shorter oligonucleotide flaps. This single nucleotide flap removal at high frequency increases the probability of ligation between the upstream and downstream DNA strands at an early phase, terminating nick translation. Besides β-clamp-mediated ligation helps DNA ligase to seal the nick promptly during the maturation of Okazaki fragments.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>29924849</pmid><doi>10.1371/journal.pone.0199559</doi><orcidid>https://orcid.org/0000-0002-0458-4109</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2018-06, Vol.13 (6), p.e0199559-e0199559 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_2062793146 |
| source | DOAJ Directory of Open Access Journals; Public Library of Science (PLoS); EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Bacillus subtilis Bacteria Biology and Life Sciences Deoxyribonucleic acid Depolymerization Displacement activity DNA DNA polymerase DNA repair DNA-directed DNA polymerase E coli Escherichia coli Exonuclease Flaps Fragments Maturation Nucleotides Okazaki fragments Oligonucleotides Physical Sciences Polypeptides Research and Analysis Methods Substrate inhibition Translation |
| title | Escherichia coli β-clamp slows down DNA polymerase I dependent nick translation while accelerating ligation |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-11T19%3A18%3A21IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Escherichia%20coli%20%CE%B2-clamp%20slows%20down%20DNA%20polymerase%20I%20dependent%20nick%20translation%20while%20accelerating%20ligation&rft.jtitle=PloS%20one&rft.au=Bhardwaj,%20Amit&rft.date=2018-06-20&rft.volume=13&rft.issue=6&rft.spage=e0199559&rft.epage=e0199559&rft.pages=e0199559-e0199559&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0199559&rft_dat=%3Cproquest_plos_%3E2057865460%3C/proquest_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2062793146&rft_id=info:pmid/29924849&rft_doaj_id=oai_doaj_org_article_240414fb3a7d4bbca4fab5f22815f941&rfr_iscdi=true |