{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T08:45:41Z","timestamp":1792140341338,"version":"4.4.0"},"reference-count":60,"publisher":"American Chemical Society (ACS)","issue":"10","license":[{"start":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T00:00:00Z","timestamp":1790899200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T00:00:00Z","timestamp":1790899200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T00:00:00Z","timestamp":1790899200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/100000165","name":"Division of Chemistry","doi-asserted-by":"publisher","award":["CHE-2529641"],"award-info":[{"award-number":["CHE-2529641"]}],"id":[{"id":"10.13039\/100000165","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Quorum sensing (QS) enables bacteria to coordinate collective behaviors through the detection of secreted signaling molecules and represents an attractive target for anti-virulence therapeutic development. In Streptococcus sinensis, an emerging opportunistic pathogen associated with infective endocarditis, QS is mediated by a competence-stimulating peptide (CSP) that activates the membrane-bound histidine kinase receptor ComD. Although previous studies demonstrated that the S. sinensis CSP is unusually tolerant to single-amino acid substitutions, the minimum structural requirements for productive receptor activation remained unknown. Here, we systematically define the molecular determinants of S. sinensis CSP activity through peptide truncation analysis, reverse alanine scanning, and inter-species crosstalk screening. Truncation studies identified the C-terminal Lys as dispensable for activity while highlighting the critical role of the N-terminus in receptor engagement. Reverse alanine scanning revealed multiple minimal functional scaffolds that retained native levels of QS activation despite replacement of nearly one-third of the peptide sequence with alanine, demonstrating that productive signaling requires only a limited subset of native residues. Consistent with these findings, screening of CSPs from diverse Streptococcus species revealed that only a small subset activated the S. sinensis ComD receptor, and only with substantially reduced potency relative to the native CSP, indicating that receptor permissiveness remains constrained by specific sequence requirements. Finally, incorporation of a D1A substitution predictably converted agonists, including minimal functional scaffolds, into competitive antagonists while preserving their relative potency trends. The most potent antagonist also demonstrated phenotypic suppression of natural competence in S. sinensis, confirming functional inhibition of QS-controlled behavior. Together, these findings define the minimal structural features required for S. sinensis QS activation, provide new insight into how receptor promiscuity is balanced with signaling specificity, and establish a foundation for the rational design of peptide-based QS modulators.<\/jats:p>","DOI":"10.1021\/acschembio.6c00715","type":"journal-article","created":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T22:01:36Z","timestamp":1790978496000},"page":"2811-2818","source":"Crossref","is-referenced-by-count":0,"title":["Minimal Functional Scaffolds of the\n                    <i>Streptococcus sinensis<\/i>\n                    Competence-Stimulating Peptide Define the Balance between Receptor Promiscuity and Signaling Specificity"],"prefix":"10.1021","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9917-274X","authenticated-orcid":true,"given":"Clay P.","family":"Renshaw","sequence":"first","affiliation":[{"name":"University of Nevada , , Reno, 1664 North Virginia Street , , ,","place":["Reno, Nevada, United States, 89557"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angela","family":"Wang","sequence":"additional","affiliation":[{"name":"University of Nevada , , Reno, 1664 North Virginia Street , , ,","place":["Reno, Nevada, United States, 89557"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher R.","family":"Yeh","sequence":"additional","affiliation":[{"name":"University of Nevada , , Reno, 1664 North Virginia Street , , ,","place":["Reno, Nevada, United States, 89557"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aspen C.","family":"Easter","sequence":"additional","affiliation":[{"name":"University of Nevada , , Reno, 1664 North Virginia Street , , ,","place":["Reno, Nevada, United States, 89557"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Umme N.","family":"Simanto","sequence":"additional","affiliation":[{"name":"University of Nevada , , Reno, 1664 North Virginia Street , , ,","place":["Reno, Nevada, United States, 89557"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2052-6782","authenticated-orcid":true,"given":"Yftah","family":"Tal-Gan","sequence":"additional","affiliation":[{"name":"University of Nevada , , Reno, 1664 North Virginia Street , , ,","place":["Reno, Nevada, United States, 89557"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,10,2]]},"reference":[{"key":"2026101523061829900_cit1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biotechadv.2025.108733","article-title":"Microbial Quorum Sensing: Mechanisms, Applications, and Challenges","volume":"86","author":"Ruan","year":"2026","journal-title":"Biotechnol. Adv."},{"issue":"1","key":"2026101523061829900_cit2","doi-asserted-by":"publisher","DOI":"10.1002\/mbo3.70016","article-title":"Quorum Sensing: Not Just a Bridge between Bacteria","volume":"14","author":"Liu","year":"2025","journal-title":"MicrobiologyOpen"},{"issue":"1","key":"2026101523061829900_cit3","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1038\/s42003-025-07608-9","article-title":"A Review of Quorum-Sensing and Its Role in Mediating Interkingdom Interactions in the Ocean","volume":"8","author":"Coolahan","year":"2025","journal-title":"Commun. Biol."},{"issue":"3","key":"2026101523061829900_cit4","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02331-17","article-title":"Bacterial Quorum Sensing and Microbial Community Interactions","volume":"9","author":"Abisado","year":"2018","journal-title":"mBio"},{"key":"2026101523061829900_cit5","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1146\/annurev.cellbio.21.012704.131001","article-title":"Quorum Sensing: Cell-to-Cell Communication in Bacteria","volume":"21","author":"Waters","year":"2005","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"2026101523061829900_cit6","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1146\/annurev.micro.55.1.165","article-title":"Quorum Sensing in Bacteria","volume":"55","author":"Miller","year":"2001","journal-title":"Annu. Rev. Microbiol."},{"issue":"5111","key":"2026101523061829900_cit7","doi-asserted-by":"publisher","first-page":"1127","DOI":"10.1126\/science.8493556","article-title":"Expression of Pseudomonas aeruginosa Virulence Genes Requires Cell-to-Cell Communication","volume":"260","author":"Passador","year":"1993","journal-title":"Science"},{"issue":"1","key":"2026101523061829900_cit8","doi-asserted-by":"publisher","first-page":"46","DOI":"10.1128\/MMBR.05007-11","article-title":"The Multiple Signaling Systems Regulating Virulence in Pseudomonas aeruginosa","volume":"76","author":"Nadal Jimenez","year":"2012","journal-title":"Microbiol. Mol. Biol. Rev."},{"issue":"6","key":"2026101523061829900_cit9","doi-asserted-by":"publisher","first-page":"2630","DOI":"10.1111\/1462-2920.15784","article-title":"Revisiting the Virulence Hallmarks of Pseudomonas aeruginosa: A Chronicle through the Perspective of Quorum Sensing","volume":"24","author":"Chadha","year":"2022","journal-title":"Environ. Microbiol."},{"issue":"1","key":"2026101523061829900_cit10","doi-asserted-by":"publisher","first-page":"461","DOI":"10.1128\/iai.00590-09","article-title":"Quorum Sensing Negatively Regulates Hemolysin Transcriptionally and Posttranslationally in Vibrio cholerae","volume":"78","author":"Tsou","year":"2010","journal-title":"Infect. Immun."},{"issue":"12","key":"2026101523061829900_cit11","doi-asserted-by":"publisher","first-page":"3794","DOI":"10.1128\/jb.186.12.3794-3805.2004","article-title":"Quorum Sensing Regulates Type III Secretion in Vibrio harveyi and Vibrio parahaemolyticus","volume":"186","author":"Henke","year":"2004","journal-title":"J. Bacteriol."},{"issue":"5","key":"2026101523061829900_cit12","doi-asserted-by":"publisher","first-page":"3129","DOI":"10.1073\/pnas.052694299","article-title":"Quorum-Sensing Regulators Control Virulence Gene Expression in Vibrio cholerae","volume":"99","author":"Zhu","year":"2002","journal-title":"Proc. Natl. Acad. Sci. U. S. 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Invest."},{"issue":"3","key":"2026101523061829900_cit18","doi-asserted-by":"publisher","first-page":"805","DOI":"10.1128\/JCM.40.3.805-810.2002","article-title":"Streptococcus sinensis sp. nov., a Novel Species Isolated from a Patient with Infective Endocarditis","volume":"40","author":"Woo","year":"2002","journal-title":"J. Clin. Microbiol."},{"issue":"10","key":"2026101523061829900_cit19","doi-asserted-by":"publisher","first-page":"2699","DOI":"10.1128\/jb.184.10.2699-2708.2002","article-title":"A Quorum-Sensing Signaling System Essential for Genetic Competence in Streptococcus mutans Is Involved in Biofilm Formation","volume":"184","author":"Li","year":"2002","journal-title":"J. Bacteriol."},{"issue":"2","key":"2026101523061829900_cit20","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1111\/j.1574-6968.2001.tb10952.x","article-title":"Effects of Mutating Putative Two-Component Systems on Biofilm Formation by Streptococcus mutans UA159","volume":"205","author":"Bhagwat","year":"2001","journal-title":"FEMS Microbiol. Lett."},{"issue":"7","key":"2026101523061829900_cit21","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2421164122","article-title":"The MutRS Quorum-Sensing System Controls Lantibiotic Mutacin Production in the Human Pathogen Streptococcus mutans","volume":"122","author":"Wyllie","year":"2025","journal-title":"Proc. Natl. Acad. Sci. U. S. 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Microbiol."},{"key":"2026101523061829900_cit27","doi-asserted-by":"publisher","first-page":"1096","DOI":"10.3390\/genes11091096","article-title":"Export of Rgg Quorum Sensing Peptides Is Mediated by the PptAB ABC Transporter in Streptococcus thermophilus Strain LMD-9","volume":"11","author":"Lingeswaran","year":"2020","journal-title":"Genes"},{"key":"2026101523061829900_cit28","doi-asserted-by":"publisher","first-page":"94","DOI":"10.3389\/fcimb.2019.00094","article-title":"Competence in Streptococcus pneumoniae and Close Commensal Relatives: Mechanisms and Implications","volume":"9","author":"Salvadori","year":"2019","journal-title":"Front. Cell. Infect. Microbiol."},{"issue":"1","key":"2026101523061829900_cit29","doi-asserted-by":"publisher","first-page":"6369","DOI":"10.1038\/s41598-018-24910-1","article-title":"Rgg-Shp Regulators Are Important for Pneumococcal Colonization and Invasion through Their Effect on Mannose Utilization and Capsule Synthesis","volume":"8","author":"Zhi","year":"2018","journal-title":"Sci. Rep."},{"issue":"5","key":"2026101523061829900_cit30","doi-asserted-by":"publisher","DOI":"10.1128\/mSphere.00324-17","article-title":"A Quorum-Sensing System That Regulates Streptococcus pneumoniae Biofilm Formation and Surface Polysaccharide Production","volume":"2","author":"Junges","year":"2017","journal-title":"mSphere"},{"issue":"1","key":"2026101523061829900_cit31","doi-asserted-by":"publisher","first-page":"15","DOI":"10.3390\/genes8010015","article-title":"Quorum Sensing Regulation of Competence and Bacteriocins in Streptococcus pneumoniae and mutans","volume":"8","author":"Shanker","year":"2017","journal-title":"Genes"},{"key":"2026101523061829900_cit32","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1146\/annurev-genet-120116-023507","article-title":"Genetic and Structural Analyses of RRNPP Intercellular Peptide Signaling of Gram-Positive Bacteria","volume":"51","author":"Neiditch","year":"2017","journal-title":"Annu. Rev. Genet."},{"issue":"2","key":"2026101523061829900_cit33","doi-asserted-by":"publisher","first-page":"181","DOI":"10.1111\/mmi.13060","article-title":"Quorum-Sensing Regulators in Gram-Positive Bacteria: \u2018Cherchez le Peptide\u2019","volume":"97","author":"Monnet","year":"2015","journal-title":"Mol. Microbiol."},{"key":"2026101523061829900_cit34","doi-asserted-by":"publisher","first-page":"1176","DOI":"10.3389\/fmicb.2015.01176","article-title":"Death and Survival in Streptococcus mutans: Differing Outcomes of a Quorum-Sensing Signaling Peptide","volume":"6","author":"Leung","year":"2015","journal-title":"Front. Microbiol."},{"issue":"1","key":"2026101523061829900_cit35","doi-asserted-by":"publisher","first-page":"105","DOI":"10.1128\/jb.00926-12","article-title":"Cell Death of Streptococcus mutans Induced by a Quorum-Sensing Peptide Occurs via a Conserved Streptococcal Autolysin","volume":"195","author":"Dufour","year":"2013","journal-title":"J. Bacteriol."},{"issue":"8","key":"2026101523061829900_cit36","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002190","article-title":"Two Group A Streptococcal Peptide Pheromones Act through Opposing Rgg Regulators to Control Biofilm Development","volume":"7","author":"Chang","year":"2011","journal-title":"PLoS Pathog."},{"issue":"6","key":"2026101523061829900_cit37","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1016\/s0923-2508(00)00171-6","article-title":"Regulation of Competence for Genetic Transformation in Streptococcus pneumoniae: A Link between Quorum Sensing and DNA Processing Genes","volume":"151","author":"Morrison","year":"2000","journal-title":"Res. Microbiol."},{"issue":"1","key":"2026101523061829900_cit38","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1046\/j.1365-2958.1998.00904.x","article-title":"Development of Competence in Streptococcus pneumoniae: Pheromone Autoinduction and Control of Quorum Sensing by the Oligopeptide Permease","volume":"29","author":"Alloing","year":"1998","journal-title":"Mol. Microbiol."},{"issue":"8","key":"2026101523061829900_cit39","doi-asserted-by":"publisher","first-page":"1270","DOI":"10.1021\/acs.biochem.6c00057","article-title":"Mapping the Structural Determinants of Quorum Sensing Activity in Streptococcus sinensis via Mutational Analysis of Its Competence Stimulating Peptide","volume":"65","author":"Renshaw","year":"2026","journal-title":"Biochemistry"},{"issue":"14","key":"2026101523061829900_cit40","doi-asserted-by":"publisher","first-page":"3104","DOI":"10.1021\/acs.biochem.5c00195","article-title":"Deciphering the Regulatory Role and Molecular Interactions That Drive the Competence Regulon Quorum Sensing Circuitry in Streptococcus constellatus","volume":"64","author":"Mehrani","year":"2025","journal-title":"Biochemistry"},{"issue":"5","key":"2026101523061829900_cit41","doi-asserted-by":"publisher","first-page":"1123","DOI":"10.1021\/acschembio.5c00180","article-title":"Elucidating the Role of the Competence Regulon Quorum Sensing Circuitry in Streptococcus cristatus","volume":"20","author":"Brennan","year":"2025","journal-title":"ACS Chem. Biol."},{"key":"2026101523061829900_cit42","doi-asserted-by":"publisher","first-page":"3631","DOI":"10.1021\/acsinfecdis.5c00926","article-title":"The Quorum Sensing-Controlled Competence Regulon Drives H2O2 Production in Streptococcus gordonii","volume":"11","author":"Brennan","year":"2025","journal-title":"ACS Infect. Dis."},{"issue":"2","key":"2026101523061829900_cit43","doi-asserted-by":"publisher","first-page":"298","DOI":"10.1016\/j.chembiol.2023.09.007","article-title":"Noncanonical Streptococcus sanguinis ComCDE Circuitry Integrates Environmental Cues in Transformation Outcome Decision","volume":"31","author":"Guo","year":"2024","journal-title":"Cell Chem. Biol."},{"key":"2026101523061829900_cit44","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.001256","article-title":"Investigating the Streptococcus sinensis Competence Regulon through a Combination of Transcriptome Analysis and Phenotypic Evaluation","volume":"168","author":"Brennan","year":"2022","journal-title":"Microbiology"},{"issue":"12","key":"2026101523061829900_cit45","doi-asserted-by":"publisher","first-page":"2834","DOI":"10.1021\/acschembio.1c00746","article-title":"Elucidating the Role and Structure-Activity Relationships of the Streptococcus oligofermentans Competence-Stimulating Peptide","volume":"16","author":"Mull","year":"2021","journal-title":"ACS Chem. Biol."},{"issue":"1","key":"2026101523061829900_cit46","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.03189-20","article-title":"Secretion, Maturation, and Activity of a Quorum Sensing Peptide (GSP) Inducing Bacteriocin Transcription in Streptococcus gallolyticus","volume":"12","author":"Harrington","year":"2021","journal-title":"mBio"},{"issue":"2","key":"2026101523061829900_cit47","doi-asserted-by":"publisher","first-page":"60","DOI":"10.1039\/d0cb00012d","article-title":"Biological Evaluation of Native Streptococcal Competence Stimulating Peptides Reveals Potential Crosstalk between Streptococcus mitis and Streptococcus pneumoniae and a New Scaffold for the Development of S. pneumoniae Quorum Sensing Modulators","volume":"1","author":"Milly","year":"2020","journal-title":"RSC Chem. Biol."},{"issue":"10","key":"2026101523061829900_cit48","doi-asserted-by":"publisher","first-page":"2833","DOI":"10.1021\/acschembio.0c00650","article-title":"Structure Activity Relationship Study of the XIP Quorum Sensing Pheromone in Streptococcus mutans Reveal Inhibitors of the Competence Regulon","volume":"15","author":"Bikash","year":"2020","journal-title":"ACS Chem. Biol."},{"key":"2026101523061829900_cit49","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioorg.2019.102987","article-title":"Exploring the Competence Stimulating Peptide (CSP) N-Terminal Requirements for Effective ComD Receptor Activation in Group1 Streptococcus pneumoniae","volume":"89","author":"Yang","year":"2019","journal-title":"Bioorg. Chem."},{"issue":"6","key":"2026101523061829900_cit50","doi-asserted-by":"publisher","first-page":"811","DOI":"10.1016\/j.bmcl.2019.01.029","article-title":"Identification of Highly Potent Competence Stimulating Peptide-Based Quorum Sensing Activators in Streptococcus mutans through the Utilization of N-Methyl and Reverse Alanine Scanning","volume":"29","author":"Bikash","year":"2019","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"2026101523061829900_cit51","doi-asserted-by":"publisher","first-page":"1769","DOI":"10.3762\/bjoc.14.151","article-title":"Defining the Hydrophobic Interactions That Drive Competence Stimulating Peptide (CSP)-ComD Binding in Streptococcus pneumoniae","volume":"14","author":"Koirala","year":"2018","journal-title":"Beilstein J. Org. Chem."},{"issue":"9","key":"2026101523061829900_cit52","doi-asserted-by":"publisher","first-page":"1385","DOI":"10.1021\/acsinfecdis.8b00115","article-title":"Structure-Activity Relationships of the Competence Stimulating Peptide in Streptococcus mutans Reveal Motifs Critical for Membrane Protease SepM Recognition and ComD Receptor Activation","volume":"4","author":"Bikash","year":"2018","journal-title":"ACS Infect. Dis."},{"issue":"4","key":"2026101523061829900_cit53","doi-asserted-by":"publisher","first-page":"1141","DOI":"10.1021\/acschembio.7b00007","article-title":"Structure-Activity Relationships of the Competence Stimulating Peptides (CSPs) in Streptococcus pneumoniae Reveal Motifs Critical for Intra-Group and Cross-Group ComD Receptor Activation","volume":"12","author":"Yang","year":"2017","journal-title":"ACS Chem. Biol."},{"issue":"6","key":"2026101523061829900_cit54","doi-asserted-by":"publisher","DOI":"10.1042\/bsr20260162","article-title":"Quorum Sensing in Streptococci and Its Peptide-Mediated Modulation","volume":"46","author":"Renshaw","year":"2026","journal-title":"Biosci. Rep."},{"issue":"5","key":"2026101523061829900_cit55","doi-asserted-by":"publisher","first-page":"299","DOI":"10.1021\/acsptsci.2c00037","article-title":"Pharmacological Evaluation of Synthetic Dominant-Negative Peptides Derived from the Competence-Stimulating Peptide of Streptococcus pneumoniae","volume":"5","author":"Oh","year":"2022","journal-title":"ACS Pharmacol. Transl. 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