Inhibition of β-lactamase-mediated oxacillin resistance in staphylococcus aureus by a deoxyribozyme

ABSTRACT AIM: To investigate the oxacillin susceptibility restoration of methicillin-resistant _Staphylococcus aureus_ (MRSA) by targeting the signaling pathway of _blaR1-blaZ_ with a

DNAzyme. METHODS: A DNAzyme (named PS-DRz602) targeting _blaR_1 mRNA was designed and synthesized. After DRz602 was introduced into a MRSA strain WHO-2, the colony-forming units of WHO-2 on

the Mueller-Hinton agar containing 6 mg/L oxacillin and the minimum inhibitory concentrations of oxacillin were determined. The inhibitory effects of DRz602 on the expressions of

antibiotic-resistant gene _blaR1_ and its downstream gene _blaZ_ were detected by real time RT-PCR. RESULTS: PS-DRz602 significantly decreased the transcription of _blaR_1 mRNA and led to

the significant reduction of _blaZ_ in a concentration-dependent manner. Consequently, the resistance of _S aureus_ WHO-2 to the β-lactam antibiotic oxacillin was significantly inhibited.

CONCLUSION: Our results indicated that blocking the _blaR1-blaZ_ signaling pathway via DNAzyme might provide a viable strategy for inhibiting the resistance of MRSA to β-lactam antibiotics

and that BlaR1 might be a potential target for pharmacological agents combating MRSA. SIMILAR CONTENT BEING VIEWED BY OTHERS RESTORING SUSCEPTIBILITY TO Β-LACTAM ANTIBIOTICS IN

METHICILLIN-RESISTANT _STAPHYLOCOCCUS AUREUS_ Article 26 July 2024 ANANDAMIDE ALTERS THE MEMBRANE PROPERTIES, HALTS THE CELL DIVISION AND PREVENTS DRUG EFFLUX IN MULTIDRUG RESISTANT

_STAPHYLOCOCCUS AUREUS_ Article Open access 22 April 2021 THE EFFECT OF NOVEL Β-LACTAM DERIVATIVES SYNTHESIZED FROM SUBSTITUTED PHENETHYLAMINES ON RESISTANCE GENES OF MRSA ISOLATES Article

29 August 2024 ARTICLE PDF REFERENCES * Aires de Sousa, M, de Lencastre H . Bridges from hospitals to the laboratory: genetic portraits of methicillin-resistant _Staphylococcus aureus_

clones. _FEMS Immunol Med Microbiol_ 2004; 40: 101–11. Article  CAS  Google Scholar  * Zetola N, Francis JS, Nuermberger EL, Bishai WR . Community-acquired methicillin-resistant

Staphylococcus aureus: an emerging threat. _Lancet Infect Dis_ 2005; 5: 275–86. Article  Google Scholar  * Vandenesch F, Naimi T, Enright MC, Lina G, Nimmo GR, Heffernan H, _et al_.

Community-acquired methicillin-resistant _Staphylococcus aureus_ carrying panton-valentine leukocidin genes: worldwide emergence. _Emerg Infect Dis_ 2003; 9: 978–84. Article  Google Scholar

  * Eady EA, Cove JH . Staphylococcal resistance revisited: community-acquired methicillin resistant _Staphylococcus aureus_ — an emerging problem for the management of skin and soft tissue

infections. _Curr Opin Infec Dis_ 2003; 16: 103–24. Article  CAS  Google Scholar  * Crum NF . The emergence of severe, community-acquired methicillin-resistant _Staphylococcus aureus_

infections. _Scand J Infec Dis_ 2005; 37: 651–6. Article  Google Scholar  * Tumbarello M, de Gaetano Donati, K, Tacconelli E, Citton R, Spanu T, Leone F, _et al_. Risk factors and predictors

of mortality of methicillin-resistant _Staphylococcus aureus_ (MRSA) bacteraemia in HIV-infected patients. _J Antimicrob Chemother_ 2002; 50: 375–82. Article  CAS  Google Scholar  *

Crowcroft NS, Catchpole M . Mortality from methicillin resistant _Staphylococcus aureus_ in England and Wales: analysis of death certificates. _BMJ_ 2002; 325: 1390–1. Article  CAS  Google

Scholar  * Srinivasan A, Dick JD, Perl TM . Vancomycin resistance in Staphylococci. _Clin Microbiol Rev_ 2002; 15: 430–8. Article  CAS  Google Scholar  * Archer GL, Bosilevac JM . Signaling

antibiotic resistance in Staphylococci. _Science_ 2001; 291: 1915–6. Article  CAS  Google Scholar  * Lowy FD . Antimicrobial resistance: the example of Staphylococcus aureus. _J Clin Invest_

2003; 111: 1265–73. Article  CAS  Google Scholar  * Safo MK, Zhao Q, Ko TP, Musayev FN, Robinson H, Scarsdale N, _et al_. Crystal structures of the BlaI repressor from _Staphylococcus

aureus_ and its complex with DNA: insights into transcriptional regulation of the bla and mec operons. _J Bacteriol_ 2005; 187: 1833–44. Article  CAS  Google Scholar  * Golemi-Kotra D, Cha

JY, Meroueh SO, Vakulenko SB, Mobashery S . Resistance to beta-lactam antibiotics and its mediation by the sensor domain of the transmembrane BlaR signaling pathway in Staphylococcus aureus.

_J Biol Chem_ 2003; 278: 18 419–25. Article  Google Scholar  * Emilsson, GM, Breaker, RR . Deoxyribozymes: new activities and new applications. _Cell Mol Life Sci_ 2002; 59: 596–607.

Article  CAS  Google Scholar  * Khachigian LM . Catalytic DNAs as potential therapeutic agents and sequence-specific molecular tools to dissect biological function. _J Clin Invest_ 2000;

106: 1189–95. Article  CAS  Google Scholar  * Bignardi GE, Woodford N, Chapman A, Johnson AP, Speller DC . Detection of the mec-A gene and phenotypic detection of resistance in

_Staphylococcus aureus_ isolates with borderline or low-level methicillin resistance. _J Antimicrob Chemother_ 1996; 37: 53–63. Article  CAS  Google Scholar  * Yazdankhah SP, Sorum H,

Oppegaard H . Comparison of genes involved in penicillin resistance in Staphylococci of bovine origin. _Microb Drug Resist_ 2000; 6: 29–36. Article  CAS  Google Scholar  * Augustin J, Gotz F

. Transformation of Staphylococcus epidermidis and other staphylococcal species with plasmid DNA by electroporation. _FEMS Microbiol Lett_ 1990; 54: 203–7. Article  CAS  Google Scholar  *

Eleaume H, Jabbouri S . Comparison of two standardisation methods in real-time quantitative RT-PCR to follow _Staphylococcus aureus_genes expression during in vitro growth. _J Microbiol

Methods_ 2004; 59: 363–70. Article  CAS  Google Scholar  * Goerke C, Bayer MG, Wolz C . Quantification of bacterial transcripts during infection using competitive reverse transcription-PCR

(RT-PCR) and LightCycler RT-PCR. _Clin Diagn Lab Immunol_ 2001; 8: 279–82. CAS  PubMed  PubMed Central  Google Scholar  * Dzidic S, Bedekovic V . Horizontal gene transfer-emerging multidrug

resistance in hospital bacteria. _Acta Pharmacol Sin_ 2003; 24: 519–26. CAS  PubMed  Google Scholar  * Johnson AP, Pearson A, Duckworth G . Surveillance and epidemiology of MRSA bacteraemia

in the UK. _J Antimicrob Chemo-ther_ 2005; 56: 455–62. Article  CAS  Google Scholar  * Chopra I . Prospects for antisense agents in the therapy of bacterial infections. _Expert Opin Investig

Drugs_ 1999; 8: 1203–8. Article  CAS  Google Scholar  * Tan XX, Actor JK, Chen Y . Peptide nucleic acid antisense oligomer as a therapeutic strategy against bacterial infection: proof of

principle using mouse intraperitoneal infection. _Antimicrob Agents Chemother_ 2005; 49: 3203–7. Article  CAS  Google Scholar  * Sarno R, Ha H, Weinsetel N, Tolmasky ME . Inhibition of

aminoglycoside 6′-N-acetyltransferase type Ib-mediated amikacin resistance by antisense oligodeoxynucleotides. _Antimicrob Agents Chemother_ 2003; 47: 3296–304. Article  CAS  Google Scholar

  * Good L, Awasthi SK, Dryselius R, Larsson O, Nielsen PE . Bactericidal antisense effects of peptide-PNA conjugates. _Nat Biotechnol_ 2001; 19: 360–4. Article  CAS  Google Scholar  *

Geller BL, Deere J, Tilley L, Iversen PL . Antisense phosphoro-diamidate morpholino oligomer inhibits viability of Escherichia coli in pure culture and in mouse peritonitis. _J Antimicrob

Chemother_ 2005; 55: 983–8. Article  CAS  Google Scholar  * White DG, Maneewannakul K, von Hofe, E, Zillman M, Eisenberg W, Field AK, _et al_. Inhibition of the multiple antibioc resistance

(mar) operon in Escherichia coli by antisense DNA analogs. _Antimicrob Agents Chemother_ 1997; 41: 2699–704. Article  CAS  Google Scholar  * Rapaport E, Levina A, Metelev V, Zamecnik PC .

Antimyco-bacterial activities of antisense oligodeoxynucleotide phosphoro-thioates in drug-resistant strains. _Proc Natl Acad Sci USA_ 1996; 93: 709–13. Article  CAS  Google Scholar  * Harth

G, Zamecnik PC, Tang JY, Tabatadze D, Horwitz MA . Treatment of Mycobaterium tuberculosis with antisense oligo-nucleotides to glutamine synthetase mRNA inhibits glutamine synthetase

activity, formation of the poly-L-glutamate/glutamine cell wall struture, and bacterial replication. _Proc Natl Acad Sci USA_ 2000; 97: 418–23. Article  CAS  Google Scholar  * Good L,

Nielsen PE . Antisense inhibition of gene expression in bacteria by PNA targeted to mRNA. _Nat Biotechnol_ 1998; 16: 355–8. Article  CAS  Google Scholar  * Torres Viera, C, Tsiodras S, Gold

HS, Coakley EP, Wennersten C, Eliopoulos GM, _et al_. Restoration of vancomycin susceptibility in Enterococcus faecalis by antiresistance determinant gene transfer. _Antimicrob Agents

Chemother_ 2001; 45: 973–5. Article  CAS  Google Scholar  * Geller BL . Antibacterial antisense. _Curr Opin Mol Ther_ 2005; 7: 109–13. CAS  PubMed  Google Scholar  * Meng JR, Hu BQ, Liu J,

Hou Z, Meng J, Jia M, _et al_. Restoration of oxacillin susceptibility in methicillin-resistant _Staphylococcus aureus_ by blocking the MecR1-mediated signaling pathway. _J Chemother_ 2006;

18: 360–5. Article  CAS  Google Scholar  * Patzel V, Steidl U, Kronenwett R, Haas R, Sczakiel G . A theoretical approach to select effective antisense oligodeoxyribonucleo-tides at high

statistical probability. _Nucleic Acids Res_ 1999; 27: 4328–34. Article  CAS  Google Scholar  * Cairns MJ, Hopkins TM, Witherington C, Wang L, Sun LQ . Target site selection for an

RNA-cleaving catalytic DNA. _Nat Biotechnol_ 1999; 17: 480–6. Article  CAS  Google Scholar  * Cairns MJ, Sun LQ . Target-site selection for the 10–23 DNAzyme. _Methods Mol Biol_ 2004; 252:

267–77. CAS  PubMed  Google Scholar  * Mitchell A, Dass CR, Sun LQ, Khachigian LM . Inhibition of human breast carcinoma proliferation, migration, chemoinvasion and solid tumour growth by

DNAzymes targeting the zinc finger transcription factor EGR-1. _Nucleic Acids Res_ 2004; 32: 3065–9. Article  CAS  Google Scholar  Download references AUTHOR INFORMATION AUTHORS AND

AFFILIATIONS * Department of Pharmacology, The Fourth Military Medical University, Xi'an, 710032, China Zheng Hou, Jing-ru Meng, Ben-quan Hu, Jie Liu, Min Jia & Xiao-xing Luo *

Institute of Genetic Diagnosis, The Fourth Military Medical University, Xi'an, 710032, China Jin-rong Zhao & Xiao-jun Yan Authors * Zheng Hou View author publications You can also

search for this author inPubMed Google Scholar * Jing-ru Meng View author publications You can also search for this author inPubMed Google Scholar * Jin-rong Zhao View author publications

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publications You can also search for this author inPubMed Google Scholar * Xiao-jun Yan View author publications You can also search for this author inPubMed Google Scholar * Min Jia View

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CORRESPONDING AUTHOR Correspondence to Xiao-xing Luo. ADDITIONAL INFORMATION This work was supported by grants from the National Natural Science Foundation of China (No 30271556) and the

Natural Science Foundation of Shaanxi Province (No 2002C2-04). RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Hou, Z., Meng, Jr., Zhao, Jr. _et al._

Inhibition of β-lactamase-mediated oxacillin resistance in _Staphylococcus aureus_ by a deoxyribozyme. _Acta Pharmacol Sin_ 28, 1775–1782 (2007).

https://doi.org/10.1111/j.1745-7254.2007.00646.x Download citation * Received: 16 January 2007 * Accepted: 17 April 2007 * Issue Date: 01 November 2007 * DOI:

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not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative KEYWORDS * methicillin-resistant _Staphylococcus aureus_ *

BlaR1 * phosphorothioate deoxyribozyme * antibiotic resistance