Please use this identifier to cite or link to this item: https://doi.org/10.3390/ijms23031661
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dc.titleThe H. pylori CagA Oncoprotein Induces DNA Double Strand Breaks through Fanconi Anemia Pathway Downregulation and Replication Fork Collapse
dc.contributor.authorKolinjivadi, Arun Mouli
dc.contributor.authorSankar, Haresh
dc.contributor.authorChoudhary, Ramveer
dc.contributor.authorTay, Lavina Sierra
dc.contributor.authorTan, Tuan Zea
dc.contributor.authorMurata-Kamiya, Naoko
dc.contributor.authorVoon, Dominic Chih-Cheng
dc.contributor.authorKappei, Dennis
dc.contributor.authorHatakeyama, Masanori
dc.contributor.authorKrishnan, Vaidehi
dc.contributor.authorIto, Yoshiaki
dc.date.accessioned2023-05-02T06:54:38Z
dc.date.available2023-05-02T06:54:38Z
dc.date.issued2022-02-01
dc.identifier.citationKolinjivadi, Arun Mouli, Sankar, Haresh, Choudhary, Ramveer, Tay, Lavina Sierra, Tan, Tuan Zea, Murata-Kamiya, Naoko, Voon, Dominic Chih-Cheng, Kappei, Dennis, Hatakeyama, Masanori, Krishnan, Vaidehi, Ito, Yoshiaki (2022-02-01). The H. pylori CagA Oncoprotein Induces DNA Double Strand Breaks through Fanconi Anemia Pathway Downregulation and Replication Fork Collapse. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 23 (3). ScholarBank@NUS Repository. https://doi.org/10.3390/ijms23031661
dc.identifier.issn1661-6596
dc.identifier.issn1422-0067
dc.identifier.urihttps://scholarbank.nus.edu.sg/handle/10635/239102
dc.description.abstractThe proteins from the Fanconi Anemia (FA) pathway of DNA repair maintain DNA replication fork integrity by preventing the unscheduled degradation of nascent DNA at regions of stalled replication forks. Here, we ask if the bacterial pathogen H. pylori exploits the fork stabilisation machinery to generate double stand breaks (DSBs) and genomic instability. Specifically, we study if the H. pylori virulence factor CagA generates host genomic DSBs through replication fork destabilisation and collapse. An inducible gastric cancer model was used to examine global CagA-dependent transcriptomic and proteomic alterations, using RNA sequencing and SILAC-based mass spectrometry, respectively. The transcriptional alterations were confirmed in gastric cancer cell lines infected with H. pylori. Functional analysis was performed using chromatin fractionation, pulsed-field gel electrophoresis (PFGE), and single molecule DNA replication/repair fiber assays. We found a core set of 31 DNA repair factors including the FA genes FANCI, FANCD2, BRCA1, and BRCA2 that were downregulated following CagA expression. H. pylori infection of gastric cancer cell lines showed downregulation of the aforementioned FA genes in a CagA-dependent manner. Consistent with FA pathway downregulation, chromatin purification studies revealed impaired levels of Rad51 but higher recruitment of the nuclease MRE11 on the chromatin of CagA-expressing cells, suggesting impaired fork protection. In line with the above data, fibre assays revealed higher fork degradation, lower fork speed, daughter strands gap accumulation, and impaired re-start of replication forks in the presence of CagA, indicating compromised genome stability. By downregulating the expression of key DNA repair genes such as FANCI, FANCD2, BRCA1, and BRCA2, H. pylori CagA compromises host replication fork stability and induces DNA DSBs through fork collapse. These data unveil an intriguing example of a bacterial virulence factor that induces genomic instability by interfering with the host replication fork stabilisation machinery.
dc.language.isoen
dc.publisherMDPI
dc.sourceElements
dc.subjectScience & Technology
dc.subjectLife Sciences & Biomedicine
dc.subjectPhysical Sciences
dc.subjectBiochemistry & Molecular Biology
dc.subjectChemistry, Multidisciplinary
dc.subjectChemistry
dc.subjectpylori
dc.subjectDNA replication
dc.subjectCagA
dc.subjectDNA double strand breaks
dc.subjectFanconi Anemia
dc.subjectgamma H2AX
dc.subjectHELICOBACTER-PYLORI
dc.subjectGASTRIC-CANCER
dc.subjectGENOME INSTABILITY
dc.subjectINFECTION
dc.subjectBRCA2
dc.subjectDEGRADATION
dc.subjectERADICATION
dc.subjectPROTEIN
dc.subjectFANCD2
dc.subjectRISK
dc.typeArticle
dc.date.updated2023-05-02T01:39:24Z
dc.contributor.departmentCANCER SCIENCE INSTITUTE OF SINGAPORE
dc.contributor.departmentMEDICINE
dc.description.doi10.3390/ijms23031661
dc.description.sourcetitleINTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
dc.description.volume23
dc.description.issue3
dc.published.statePublished
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