{"doi":"10.1128/mbio.01844-24","title":"m\n            <sup>6</sup>\n            A demethylase CpALKBH regulates\n            <i>CpZap1</i>\n            mRNA stability to modulate the development and virulence of chestnut blight fungus","abstract":"<jats:title>ABSTRACT</jats:title>\n          <jats:sec>\n            <jats:title/>\n            <jats:p>\n              As the most abundant eukaryotic mRNA modification, N\n              <jats:sup>6</jats:sup>\n              -methyladenosine (m\n              <jats:sup>6</jats:sup>\n              A) plays a crucial role in regulating multiple biological processes. This methylation is regulated by methyltransferases and demethylases. However, the regulatory role and mode of action of m\n              <jats:sup>6</jats:sup>\n              A demethylases in fungi remain poorly understood. In this study, we demonstrate that CpALKBH is a demethylase in\n              <jats:italic>Cryphonectria parasitica</jats:italic>\n              that removes m\n              <jats:sup>6</jats:sup>\n              A modification from single-stranded RNA\n              <jats:italic>in vitro</jats:italic>\n              . The deletion of\n              <jats:italic>CpALKBH</jats:italic>\n              resulted in a significant increase in the m\n              <jats:sup>6</jats:sup>\n              A methylation levels, along with decreases in the growth rate, sporulation, and virulence in\n              <jats:italic>C. parasitica</jats:italic>\n              . Additionally, CpZap1—a transcription factor—was identified as a downstream target of CpALKBH demethylase based on RNA sequencing analysis. We confirmed that CpALKBH demethylase regulates\n              <jats:italic>CpZap1</jats:italic>\n              mRNA stability in an m\n              <jats:sup>6</jats:sup>\n              A-dependent manner. Furthermore, through MazF assay, we found that methylation of\n              <jats:italic>CpZap1</jats:italic>\n              at position 1935A is regulated by both CpALKBH demethylase and CpMTA1 methyltransferase.\n              <jats:italic>CpZap1</jats:italic>\n              significantly influences the fungal phenotype and virulence, thereby restoring the abnormal phenotype observed in ∆\n              <jats:italic>CpALKBH</jats:italic>\n              mutants. Collectively, our findings highlight the essential role of CpALKBH as an m\n              <jats:sup>6</jats:sup>\n              A demethylase in the development and virulence of\n              <jats:italic>C. parasitica</jats:italic>\n              , while also elucidating the molecular mechanisms through which m\n              <jats:sup>6</jats:sup>\n              A modification impacts\n              <jats:italic>CpZap1</jats:italic>\n              mRNA stability.\n            </jats:p>\n          </jats:sec>\n          <jats:sec>\n            <jats:title>IMPORTANCE</jats:title>\n            <jats:p>\n              N\n              <jats:sup>6</jats:sup>\n              -methyladenosine (m\n              <jats:sup>6</jats:sup>\n              A) is the most abundant eukaryotic mRNA modification and is involved in various biological processes. Methyltransferases and demethylases regulate the m\n              <jats:sup>6</jats:sup>\n              A modification, but the regulatory role of m\n              <jats:sup>6</jats:sup>\n              A demethylases in fungi remains poorly understood. Here, we demonstrated that CpALKBH functions as a demethylase in\n              <jats:italic>Cryphonectria parasitica</jats:italic>\n              . The deletion of\n              <jats:italic>CpALKBH</jats:italic>\n              leads to a significant increase in m\n              <jats:sup>6</jats:sup>\n              A levels and a reduction in fungal growth, sporulation, and virulence. We identified\n              <jats:italic>CpZap1</jats:italic>\n              as a downstream target of CpALKBH, with CpALKBH regulating\n              <jats:italic>CpZap1</jats:italic>\n              mRNA stability in an m\n              <jats:sup>6</jats:sup>\n              A-dependent manner. Additionally, our findings indicate that methylation at position 1935A of\n              <jats:italic>CpZap1</jats:italic>\n              is regulated by both the CpALKBH demethylase and the CpMTA1 methyltransferase. Given its critical role in fungal development and virulence, overexpression of\n              <jats:italic>CpZap1</jats:italic>\n              can rescue abnormal phenotypes of ∆\n              <jats:italic>CpALKBH</jats:italic>\n              mutant. Overall, these findings contribute to improving our understanding of the role of m\n              <jats:sup>6</jats:sup>\n              A demethylase in fungi.\n            </jats:p>\n          </jats:sec>","journal":"mBio","year":2025,"id":617790,"datarank":0.26592723806066304,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"self_citation_contribution":0.24141568686511508,"citation_network_contribution":0.024511551195547973,"self_endowment_contribution":0.24141568686511508,"citer_contribution":0.024511551195547973,"corpus_percentile":null,"corpus_rank":null,"citation_count":4,"citer_count":4,"citers_with_citation_signal":2,"citers_with_endowment":2,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1386061,"name":"Xiangyu Wei","orcid":"0000-0002-8774-5296","position":1,"is_corresponding":false},{"id":1593349,"name":"Fengyue Chen","orcid":null,"position":2,"is_corresponding":false},{"id":378425,"name":"Baoshan Chen","orcid":"0000-0002-7001-215X","position":3,"is_corresponding":false},{"id":769145,"name":"Ru Li","orcid":"0000-0002-9756-793X","position":4,"is_corresponding":false},{"id":1593348,"name":"Lijiu Zhao","orcid":"0009-0009-7097-4031","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"m\n            <sup>6</sup>\n            A demethylase CpALKBH regulates\n            <i>CpZap1</i>\n            mRNA stability to modulate the development and virulence of chestnut blight fungus","abstract":"<jats:title>ABSTRACT</jats:title>\n          <jats:sec>\n            <jats:title/>\n            <jats:p>\n              As the most abundant eukaryotic mRNA modification, N\n              <jats:sup>6</jats:sup>\n              -methyladenosine (m\n              <jats:sup>6</jats:sup>\n              A) plays a crucial role in regulating multiple biological processes. This methylation is regulated by methyltransferases and demethylases. However, the regulatory role and mode of action of m\n              <jats:sup>6</jats:sup>\n              A demethylases in fungi remain poorly understood. In this study, we demonstrate that CpALKBH is a demethylase in\n              <jats:italic>Cryphonectria parasitica</jats:italic>\n              that removes m\n              <jats:sup>6</jats:sup>\n              A modification from single-stranded RNA\n              <jats:italic>in vitro</jats:italic>\n              . The deletion of\n              <jats:italic>CpALKBH</jats:italic>\n              resulted in a significant increase in the m\n              <jats:sup>6</jats:sup>\n              A methylation levels, along with decreases in the growth rate, sporulation, and virulence in\n              <jats:italic>C. parasitica</jats:italic>\n              . Additionally, CpZap1—a transcription factor—was identified as a downstream target of CpALKBH demethylase based on RNA sequencing analysis. We confirmed that CpALKBH demethylase regulates\n              <jats:italic>CpZap1</jats:italic>\n              mRNA stability in an m\n              <jats:sup>6</jats:sup>\n              A-dependent manner. Furthermore, through MazF assay, we found that methylation of\n              <jats:italic>CpZap1</jats:italic>\n              at position 1935A is regulated by both CpALKBH demethylase and CpMTA1 methyltransferase.\n              <jats:italic>CpZap1</jats:italic>\n              significantly influences the fungal phenotype and virulence, thereby restoring the abnormal phenotype observed in ∆\n              <jats:italic>CpALKBH</jats:italic>\n              mutants. Collectively, our findings highlight the essential role of CpALKBH as an m\n              <jats:sup>6</jats:sup>\n              A demethylase in the development and virulence of\n              <jats:italic>C. parasitica</jats:italic>\n              , while also elucidating the molecular mechanisms through which m\n              <jats:sup>6</jats:sup>\n              A modification impacts\n              <jats:italic>CpZap1</jats:italic>\n              mRNA stability.\n            </jats:p>\n          </jats:sec>\n          <jats:sec>\n            <jats:title>IMPORTANCE</jats:title>\n            <jats:p>\n              N\n              <jats:sup>6</jats:sup>\n              -methyladenosine (m\n              <jats:sup>6</jats:sup>\n              A) is the most abundant eukaryotic mRNA modification and is involved in various biological processes. Methyltransferases and demethylases regulate the m\n              <jats:sup>6</jats:sup>\n              A modification, but the regulatory role of m\n              <jats:sup>6</jats:sup>\n              A demethylases in fungi remains poorly understood. Here, we demonstrated that CpALKBH functions as a demethylase in\n              <jats:italic>Cryphonectria parasitica</jats:italic>\n              . The deletion of\n              <jats:italic>CpALKBH</jats:italic>\n              leads to a significant increase in m\n              <jats:sup>6</jats:sup>\n              A levels and a reduction in fungal growth, sporulation, and virulence. We identified\n              <jats:italic>CpZap1</jats:italic>\n              as a downstream target of CpALKBH, with CpALKBH regulating\n              <jats:italic>CpZap1</jats:italic>\n              mRNA stability in an m\n              <jats:sup>6</jats:sup>\n              A-dependent manner. Additionally, our findings indicate that methylation at position 1935A of\n              <jats:italic>CpZap1</jats:italic>\n              is regulated by both the CpALKBH demethylase and the CpMTA1 methyltransferase. Given its critical role in fungal development and virulence, overexpression of\n              <jats:italic>CpZap1</jats:italic>\n              can rescue abnormal phenotypes of ∆\n              <jats:italic>CpALKBH</jats:italic>\n              mutant. Overall, these findings contribute to improving our understanding of the role of m\n              <jats:sup>6</jats:sup>\n              A demethylase in fungi.\n            </jats:p>\n          </jats:sec>","is_dataset_classified":null,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"39611846","pmcid":"PMC11708048","openalex_id":"https://openalex.org/W4404851488","authors":[],"funders":[{"funder_name":"MOST | National Natural Science Foundation of China","grant_id":"32160623","title":null},{"funder_name":"Innovation Project of Guangxi Graduate Education","grant_id":"YCBZ2022032","title":null}],"total_grants":2,"fwci":0.6467,"citation_percentile":0.67372815,"influential_citations":0,"citation_trend":[{"year":2025,"count":2},{"year":2026,"count":2}],"oa_status":"gold","license":"cc-by","oa_locations":[{"url":"https://doi.org/10.1128/mbio.01844-24","host_type":"journal"},{"url":"https://doi.org/10.1128/mbio.01844-24","host_type":"publisher"},{"url":"https://journals.asm.org/doi/pdf/10.1128/mbio.01844-24","host_type":"publisher"},{"url":"https://pubmed.ncbi.nlm.nih.gov/39611846","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/11708048","host_type":"repository"},{"url":"https://doaj.org/article/97da6f483de34b219fc411448bfcfbf6","host_type":"repository"},{"url":"https://pmc.ncbi.nlm.nih.gov/articles/PMC11708048/pdf/mbio.01844-24.pdf","host_type":"repository"},{"url":"https://europepmc.org/articles/PMC11708048","host_type":"Europe_PMC"},{"url":"https://europepmc.org/articles/PMC11708048?pdf=render","host_type":"Europe_PMC"}],"fields_of_study":["RNA modifications and cancer","Plant Disease Resistance and Genetics","Plant and Fungal Interactions Research","Virulence","RNA Stability","Ascomycota","Gene Expression Regulation, Fungal","Adenosine","Plant Diseases","Fungal Proteins","Methylation","RNA, Messenger"],"mesh_terms":["Adenosine","Ascomycota","Fungal Proteins","Methylation","Plant Diseases","RNA, Messenger","Virulence","Gene Expression Regulation, Fungal","RNA Stability"],"keywords":["Demethylase","Methyltransferase","Biology","Methylation","N6-Methyladenosine","Virulence","Messenger RNA","RNA","RNA methylation","Mutant","Cell biology","Epigenetics","Biochemistry","Gene","Cryphonectria parasitica","M6a Demethylase","Cpalkbh","Cpzap1"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Zero hunger"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"gen"},{"name":"doi"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-03T02:38:11.423514Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}