{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T12:11:21Z","timestamp":1785413481217,"version":"3.56.0"},"reference-count":37,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T00:00:00Z","timestamp":1782172800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T00:00:00Z","timestamp":1782172800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000072","name":"National Institute of Dental and Craniofacial Research","doi-asserted-by":"publisher","award":["R01DE022350"],"award-info":[{"award-number":["R01DE022350"]}],"id":[{"id":"10.13039\/100000072","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mSystems"],"published-print":{"date-parts":[[2026,6,23]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      In many gram-positive bacteria, the diadenylate cyclase DacA synthesizes the essential second messenger c-di-AMP. While regulators like GlmM and CdaR are known, the broader DacA interactome and its functional relevance remain poorly defined. Using\n                      <jats:italic toggle=\"yes\">Streptococcus mutans<\/jats:italic>\n                      as a model, we identified novel protein-protein interactions using co-immunoprecipitation and mass spectrometry approaches. We identified 22 candidate interactors under non-crosslinked conditions and 18 under crosslinked conditions, with six shared candidates. Using AlphaFold-guided modeling and experimental split-luciferase complementation, we validated SMU_723\u2014a putative calcium transporter\u2014as a primary binding partner. Targeted mutagenesis identified threonine 147, glutamine 148, and threonine 149 as the key interacting interface; mutations in these residues impaired binding, confirming their structural importance. Disruption of the DacA\u2013SMU_723 interaction resulted in a significant reduction in intracellular c-di-AMP levels, supporting a role for SMU_723 in modulating DacA enzymatic activity. Deletion of SMU_723 recapitulated several phenotypes observed in the \u2206\n                      <jats:italic toggle=\"yes\">dacA<\/jats:italic>\n                      strain, including an extended lag phase, altered cell morphology, reduced acid production and acid tolerance, impaired sorbitol metabolism, decreased colonization in the\n                      <jats:italic toggle=\"yes\">Drosophila<\/jats:italic>\n                      model, and delayed growth following calcium stimulation. However, \u2206SMU_723 phenotypes were generally less severe than those of \u2206\n                      <jats:italic toggle=\"yes\">dacA<\/jats:italic>\n                      , indicating partial functional overlap rather than a complete phenocopy. Together, this study identifies a functional link between SMU_723 and DacA that modulates c-di-AMP levels and calcium response and is vital for the survival of\n                      <jats:italic toggle=\"yes\">S. mutans<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      Mapping the DacA interactome reveals how environmental and intracellular cues tune c-di-AMP signaling to control stress adaptation, ion balance, and virulence traits in\n                      <jats:italic toggle=\"yes\">Streptococcus mutans<\/jats:italic>\n                      . By identifying new DacA-associated proteins and validating SMU_723 as a previously unrecognized interactor with genetic and phenotypic linkage to DacA, this study provides new insights into the mechanistic framework for c-di-AMP regulation in\n                      <jats:italic toggle=\"yes\">S. mutans<\/jats:italic>\n                      . The connection between DacA and a putative calcium transporter highlights a plausible axis that couples second-messenger signaling to calcium homeostasis, with implications for biofilm physiology and pathogenesis. These insights open new avenues to therapeutically modulate c-di-AMP pathways.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/msystems.01727-25","type":"journal-article","created":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T13:00:11Z","timestamp":1780491611000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Decoding the interactome for cyclic-di-AMP-producing enzyme diadenylate cyclase"],"prefix":"10.1128","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6846-3659","authenticated-orcid":false,"given":"Rong","family":"Mu","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/009avj582","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biomaterial and Biomedical Sciences, School of Dentistry, Oregon Health and Science University","place":["Portland, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baotong","family":"Xie","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/009avj582","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biomaterial and Biomedical Sciences, School of Dentistry, Oregon Health and Science University","place":["Portland, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0883-104X","authenticated-orcid":true,"given":"Stephanie","family":"Momeni","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/009avj582","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biomaterial and Biomedical Sciences, School of Dentistry, Oregon Health and Science University","place":["Portland, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1694-3954","authenticated-orcid":true,"given":"Hui","family":"Wu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/009avj582","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biomaterial and Biomedical Sciences, School of Dentistry, Oregon Health and Science University","place":["Portland, USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0086096"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00462-18"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.15187"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00564-15"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.00023-23"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1016-8478(23)13181-5"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00769-13"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41579-023-00865-7"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00147-19"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.gpp3-0007-2018"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1007342"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.GPP3-0051-2018"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.13440"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.13123"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens9050374"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.00614"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00597-18"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2021.101317"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.15008"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.13281"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M117.818716"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms8091269"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07487-w"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4792"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2021.167015"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1126\/scisignal.aal3011"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002217"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1111\/mmi.13456"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.1126\/scisignal.aaf7279"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.RA119.008774"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.27.2.444-448.1980"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.02150"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.12953"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.01500-23"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2023.1279380"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.3791\/55024"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1128\/msphere.00778-23"}],"container-title":["mSystems"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/msystems.01727-25","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/msystems.01727-25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T13:03:58Z","timestamp":1783515838000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/msystems.01727-25"}},"subtitle":[],"editor":[{"given":"Egon Anderson","family":"Ozer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,6,23]]},"references-count":37,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2026,6,23]]}},"alternative-id":["10.1128\/msystems.01727-25"],"URL":"https:\/\/doi.org\/10.1128\/msystems.01727-25","relation":{},"ISSN":["2379-5077"],"issn-type":[{"value":"2379-5077","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,23]]},"assertion":[{"value":"2025-12-10","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-03-24","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-06-03","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e01727-25"}}