The Role of Restriction-Modification Systems in the Horizontal Transfer of Carbapenem Resistance Genes in Klebsiella pneumoniae

The Role of Restriction-Modification Systems in the Horizontal Transfer of Carbapenem Resistance Genes in Klebsiella pneumoniae | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article The Role of Restriction-Modification Systems in the Horizontal Transfer of Carbapenem Resistance Genes in Klebsiella pneumoniae Huimin Chen, Qianqian Fang, Lin Zheng, Wei Liang This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7168145/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The rising detection rates of carbapenem-resistant Klebsiella pneumoniae (CRKP) pose significant challenges to clinical anti-infection treatments. The horizontal transfer of antibiotic resistance genes (ARGs) via plasmids is one of the primary mechanisms of acquired bacterial resistance. However, data analysis regarding the genetic mobility of these resistance genes remains relatively scarce. In this study, we analyzed the genomic sequences of 428 K. pneumoniae strains from the GenBank database over the past three years, focusing on Restriction-Modification (RM) system typing, plasmid typing, and resistance gene assessment. Our findings revealed a consistent distribution of RM systems among the 428 K. pneumoniae strains across China and other countries, with Type II RM systems being predominant (98.7%). These systems recognize the methylation and restriction site CCWGG (W = A or T). In other clinically common carbapenem-resistant bacteria, such as Acinetobacter baumannii and Pseudomonas aeruginosa , the proportion of Type II RM systems is only 2%. However, in K. pneumoniae and Escherichia coli strains, the carriage rate of M.EcoRII methyltransferase is 100%. From the perspective of bacterial RM defense and the horizontal transfer of mobile genetic elements (MGEs), the methylation and restriction enzyme recognition sites in K. pneumoniae are characteristic of Type II RM systems, sharing the same recognition sequence CCWGG. This may suggests that the corresponding methylated MGEs in K. pneumoniae not only spread horizontally within the species but can also engage in gene exchange with carbapenem-resistant Escherichia coli (CREC) in clinical settings. There is no difference in the distribution types of RM systems between carbapenem-resistant and sensitive K. pneumoniae . However, further analysis of carbapenem-resistant plasmids shows that the prevalence of RM systems is 33.15%. Plasmids play a central role in the epidemiology of antibiotic resistance, and the increased prevalence of RM systems in resistant gene plasmids provides more opportunities for the horizontal transfer of MGEs. In K. pneumoniae , the bla KPC−2 gene is primarily carried by IncF plasmids and is associated with multiple sequence types. However, compared to IncF, IncN, and IncH, IncA/C plasmids possess a more diverse repertoire of carbapenem ARGs. The findings of this study emphasize the key role of RM systems and plasmids in the spread of CRKP. They provide epigenetic insights into the horizontal gene transfer (HGT) of K. pneumoniae , thereby offering potential strategies for controlling the spread of ARGs. Carbapenem-resistant K. pneumoniae RM system Horizontal gene transfer Mobile genetic elements Full Text Additional Declarations No competing interests reported. Supplementary Files datasetS1.xlsx datasetS2.xlsx datasetS3.xlsx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7168145","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":502091212,"identity":"05c942a5-3e37-4962-ae71-68a033ac100c","order_by":0,"name":"Huimin Chen","email":"","orcid":"","institution":"The First Affiliated Hospital of Ningbo University","correspondingAuthor":false,"prefix":"","firstName":"Huimin","middleName":"","lastName":"Chen","suffix":""},{"id":502091213,"identity":"472dcd13-1940-462f-a34d-6e9e4222d5d2","order_by":1,"name":"Qianqian Fang","email":"","orcid":"","institution":"The First Affiliated Hospital of Ningbo 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The horizontal transfer of antibiotic resistance genes (ARGs) via plasmids is one of the primary mechanisms of acquired bacterial resistance. However, data analysis regarding the genetic mobility of these resistance genes remains relatively scarce. In this study, we analyzed the genomic sequences of 428 \u003cem\u003eK. pneumoniae\u003c/em\u003e strains from the GenBank database over the past three years, focusing on Restriction-Modification (RM) system typing, plasmid typing, and resistance gene assessment. Our findings revealed a consistent distribution of RM systems among the 428 \u003cem\u003eK. pneumoniae\u003c/em\u003e strains across China and other countries, with Type II RM systems being predominant (98.7%). These systems recognize the methylation and restriction site CCWGG (W\u0026thinsp;=\u0026thinsp;A or T). In other clinically common carbapenem-resistant bacteria, such as \u003cem\u003eAcinetobacter baumannii\u003c/em\u003e and \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e, the proportion of Type II RM systems is only 2%. However, in \u003cem\u003eK. pneumoniae\u003c/em\u003e and \u003cem\u003eEscherichia coli\u003c/em\u003e strains, the carriage rate of M.EcoRII methyltransferase is 100%. From the perspective of bacterial RM defense and the horizontal transfer of mobile genetic elements (MGEs), the methylation and restriction enzyme recognition sites in \u003cem\u003eK. pneumoniae\u003c/em\u003e are characteristic of Type II RM systems, sharing the same recognition sequence CCWGG. This may suggests that the corresponding methylated MGEs in \u003cem\u003eK. pneumoniae\u003c/em\u003e not only spread horizontally within the species but can also engage in gene exchange with carbapenem-resistant \u003cem\u003eEscherichia coli\u003c/em\u003e (CREC) in clinical settings. 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