Structural and biochemical characterization of MepR, a multidrug binding transcription regulator of the Staphylococcus aureus multidrug efflux pump MepA

Kumaraswami, Muthiah; Schuman, Jason T.; Seo, Susan M.; Kaatz, Glenn W.; Brennan, Richard G.

doi:10.1093/nar/gkn1046

Title: Structural and biochemical characterization of MepR, a multidrug binding transcription regulator of the Staphylococcus aureus multidrug efflux pump MepA

Journal Article · Wed Jan 07 00:00:00 EST 2009 · Nucleic Acids Research

DOI:https://doi.org/10.1093/nar/gkn1046· OSTI ID:1625442

Kumaraswami, Muthiah ^[1]; Schuman, Jason T. ^[2]; Seo, Susan M. ^[3]; Kaatz, Glenn W. ^[4]; Brennan, Richard G. ^[1]

Univ. of Texas, Houston, TX (United States). M.D. Anderson Cancer Center. Dept. of Biochemistry and Molecular Biology
GE Healthcare, Piscataway, NJ (United States); Oregon Health and Science Univ., Portland, OR (United States). Dept. of Biochemistry and Molecular Biology
Wayne State Univ., Detroit, MI (United States). School of Medicine. Division of Infectious Diseases
Wayne State Univ., Detroit, MI (United States). School of Medicine. John D. Dingell Veterans Administration Medical Center

MepR is a multidrug binding transcription regulator that represses expression of the Staphylococcus aureus multidrug efflux pump gene, mepA, as well as its own gene. MepR is induced by multiple cationic toxins, which are also substrates of MepA. In order to understand the gene regulatory and drug-binding mechanisms of MepR, we carried out biochemical, in vivo and structural studies. The 2.40 A' resolution structure of drug-free MepR reveals the most open MarR family protein conformation to date, which will require a huge conformational change to bind cognate DNA. DNA-binding data show that MepR uses a dual regulatory binding mode as the repressor binds the mepA operator as a dimer of dimers, but binds the mepR operator as a single dimer. Alignment of the six half sites reveals the consensus MepR binding site, 5’-GTTAGAT-3’. ‘Drug’ binding studies show that MepR binds to ethidium and DAPI with comparable affinities (K_d = 2.6 and 4.5 μM, respectively), but with significantly lower affinity to the larger rhodamine 6G (K_d = 62.6 μM). Mapping clinically relevant or in vitro selected MepR mutants onto the MepR structure suggests that their defective repressor phenotypes are due to structural and allosteric defects.

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Research Organization:: Univ. of Texas, Houston, TX (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division

Grant/Contract Number:: AC03-76SF00098

OSTI ID:: 1625442

Journal Information:: Nucleic Acids Research, Vol. 37, Issue 4; ISSN 0305-1048

Publisher:: Oxford University PressCopyright Statement

Country of Publication:: United States

Language:: English

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