Biochemical and structural analyses reveal critical residues in δ subunit affecting its bindings to β′ subunit of Staphylococcus aureus RNA polymerase

2021 ◽  
Vol 545 ◽  
pp. 98-104
Author(s):  
Zhaozhu Lin ◽  
Fulin Wang ◽  
Zhuo Shang ◽  
Wei Lin
Keyword(s):  
Staphylococcus Aureus ◽  
Rna Polymerase ◽  
Β Subunit ◽  
Critical Residues

scholarly journals A Mutation of RNA Polymerase β′ Subunit (RpoC) Converts Heterogeneously Vancomycin-Intermediate Staphylococcus aureus (hVISA) into “Slow VISA”

2015 ◽  
Vol 59 (7) ◽  
pp. 4215-4225 ◽  
Author(s):  
Miki Matsuo ◽  
Tomomi Hishinuma ◽  
Yuki Katayama ◽  
Keiichi Hiramatsu
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Rna Polymerase ◽  
Stringent Response ◽  
Whole Genome ◽  
Multicopy Plasmid ◽  
Β Subunit ◽  
Whole Genome Analysis ◽  
Content Type ◽  
Peptidoglycan Biosynthesis

ABSTRACTVarious mutations in therpoBgene, which encodes the RNA polymerase β subunit, are associated with increased vancomycin (VAN) resistance in vancomycin-intermediateStaphylococcus aureus(VISA) and heterogeneously VISA (hVISA) strains. We reported thatrpoBmutations are also linked to the expression of the recently found “slow VISA” (sVISA) phenotype (M. Saito, Y. Katayama, T. Hishinuma, A. Iwamoto, Y. Aiba, K Kuwahara-Arai, L. Cui, M. Matsuo, N. Aritaka, and K. Hiramatsu, Antimicrob Agents Chemother 58:5024–5035, 2014,http://dx.doi.org/10.1128/AAC.02470-13). Because RpoC and RpoB are components of RNA polymerase, we examined the effect of therpoC(P440L) mutation on the expression of the sVISA phenotype in the Mu3fdh2*V6-5 strain (V6-5), which was derived from a previously reported hVISA strain with the VISA phenotype. V6-5 had an extremely prolonged doubling time (DT) (72 min) and high vancomycin MIC (16 mg/liter). However, the phenotype of V6-5 was unstable, and the strain frequently reverted to hVISA with concomitant loss of low growth rate, cell wall thickness, and reduced autolysis. Whole-genome sequencing of phenotypic revertant strain V6-5-L1 and comparison with V6-5 revealed a second mutation, F562L, inrpoC. Introduction of the wild-type (WT)rpoCgene using a multicopy plasmid resolved the sVISA phenotype of V6-5, indicating that therpoC(P440L) mutant expressed the sVISA phenotype in hVISA. To investigate the mechanisms of resistance in the sVISA strain, we independently isolated an additional 10 revertants to hVISA and VISA. In subsequent whole-genome analysis, we identified compensatory mutations in the genes of three distinct functional categories: therpoCgene itself as regulatory mutations, peptidoglycan biosynthesis genes, andrelQ, which is involved in the stringent response. It appears that therpoC(P440L) mutation causes the sVISA phenotype by augmenting cell wall peptidoglycan synthesis and through the control of the stringent response.

scholarly journals RNA Polymerase Inhibitors with Activity against Rifampin-Resistant Mutants of Staphylococcus aureus

2000 ◽  
Vol 44 (11) ◽  
pp. 3163-3166 ◽  
Author(s):  
Alexander O'Neill ◽  
Brunello Oliva ◽  
Christopher Storey ◽  
Anthony Hoyle ◽  
Colin Fishwick ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Drug Development ◽  
Rna Polymerase ◽  
Gene Mutations ◽  
Β Subunit ◽  
Polymerase Inhibitors ◽  
Resistant Mutants ◽  
Sorangicin A

ABSTRACT A collection of rifampin-resistant mutants of Staphylococcus aureus with characterized RNA polymerase β-subunit (rpoB) gene mutations was cross-screened against a number of other RNA polymerase inhibitors to correlate susceptibility with specific rpoB genotypes. The rpoB mutants were cross-resistant to streptolydigin and sorangicin A. In contrast, thiolutin, holomycin, corallopyronin A, and ripostatin A retained activity against the rpoB mutants. The second group of inhibitors may be of interest as drug development candidates.

scholarly journals Mutation of RNA Polymerase β-Subunit Gene Promotes Heterogeneous-to-Homogeneous Conversion of β-Lactam Resistance in Methicillin-Resistant Staphylococcus aureus

2013 ◽  
Vol 57 (10) ◽  
pp. 4861-4871 ◽  
Author(s):  
Yoshifumi Aiba ◽  
Yuki Katayama ◽  
Tomomi Hishinuma ◽  
Hiroko Murakami-Kuroda ◽  
Longzhu Cui ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Rna Polymerase ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Phenotypic Expression ◽  
Amino Acid Replacement ◽  
Whole Genome ◽  
Β Subunit ◽  
Methicillin Resistant ◽  
Content Type

ABSTRACTThree types of phenotypic expression of β-lactam resistance have been reported in methicillin-resistantStaphylococcus aureus(MRSA): heterogeneous, homogeneous, and Eagle-type resistance. Heterogeneous-to-homogeneous conversion of β-lactam resistance is postulated to be caused by a chromosomal mutation (chr*) in addition to the expression of themecAgene. Eagle-type resistance is a unique phenotype ofchr*occurring in pre-MRSA strain N315 whosemecAgene expression is strongly repressed by an intactmecIgene. We here report that certain mutations of therpoBgene, encoding the RNA polymerase β subunit, belong tochr*. We studied homogeneous MRSA (homo-MRSA) strain N315ΔIP-H5 (abbreviated as ΔIP-H5), which was obtained from hetero-MRSA strain N315ΔIP by selection with 8 mg/liter imipenem. Whole-genome sequencing of ΔIP-H5 revealed the presence of a unique mutation in therpoBgene,rpoB(N967I), causing the amino acid replacement of Asn by Ile at position 967 of RpoB. The effect of therpoB(N967I) mutation was confirmed by constructing a revertant H5rpoB(I967N) strain as well as an N315-derived mutant, N315rpoB(N967I). H5rpoB(I967N) regained the hetero-resistance phenotype, and the N315rpoB(N967I) strain showed an Eagle-type phenotype similar to that of the typical Eagle-type MRSA strain N315h4. Furthermore, subsequent whole-genome sequencing revealed that N315h4 also had a missense mutation ofrpoB(R644H). Introduction of therpoB(N967I) mutation was accompanied by decreased autolysis, prolonged doubling time, and tolerance to bactericidal concentrations of methicillin. We consider thatrpoBmutations are the major cause for heterogeneous-to-homogeneous phenotypic conversion of β-lactam resistance in MRSA strain N315 and its derived strains.

scholarly journals CBR antimicrobials alter coupling between the bridge helix and the β subunit in RNA polymerase

2014 ◽  
Vol 5 (1) ◽  
Author(s):  
Anssi M. Malinen ◽  
Monali NandyMazumdar ◽  
Matti Turtola ◽  
Henri Malmi ◽  
Thadee Grocholski ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Β Subunit

scholarly journals Nudeotide sequence at the end of the gene for the RNA polymerase β subunit (rpoC)

1981 ◽  
Vol 9 (24) ◽  
pp. 6827-6840 ◽  
Author(s):  
C. Squires ◽  
A. Krainer ◽  
G. Barry ◽  
W.-F. Shen ◽  
C.L. Squires
Keyword(s):  
Rna Polymerase ◽  
Β Subunit

A mutation in the RNA polymerase β′ subunit causing depressed ribosomal RNA synthesis in Escherichia coli

1981 ◽  
Vol 183 (1) ◽  
pp. 54-58 ◽  
Author(s):  
Ben A. Oostra ◽  
Klaas Kok ◽  
Adri J. Van Vliet ◽  
AB Geert ◽  
Max Gruber
Keyword(s):  
Escherichia Coli ◽  
Rna Polymerase ◽  
Ribosomal Rna ◽  
Rna Synthesis ◽  
Β Subunit

Genetic studies on the β subunit of Escherichia coli RNA polymerase

1986 ◽  
Vol 203 (2) ◽  
pp. 265-268 ◽  
Author(s):  
Robert E. Glass ◽  
Steven T. Jones ◽  
Akira Ishihama
Keyword(s):  
Escherichia Coli ◽  
Rna Polymerase ◽  
Β Subunit ◽  
Genetic Studies

scholarly journals In Vitro Evaluation of CBR-2092, a Novel Rifamycin-Quinolone Hybrid Antibiotic: Studies of the Mode of Action in Staphylococcus aureus

2008 ◽  
Vol 52 (7) ◽  
pp. 2313-2323 ◽  
Author(s):  
Gregory T. Robertson ◽  
Eric J. Bonventre ◽  
Timothy B. Doyle ◽  
Qun Du ◽  
Leonard Duncan ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Rna Polymerase ◽  
Mode Of Action ◽  
Bacterial Infections ◽  
Dna Gyrase ◽  
Topoisomerase Iv ◽  
Dna Topoisomerase ◽  
Resistant Variant ◽  
Proven Efficacy

ABSTRACT Rifamycins have proven efficacy in the treatment of persistent bacterial infections. However, the frequency with which bacteria develop resistance to rifamycin agents restricts their clinical use to antibiotic combination regimens. In a program directed toward the synthesis of rifamycins with a lower propensity to elicit resistance development, a series of compounds were prepared that covalently combine rifamycin and quinolone pharmacophores to form stable hybrid antibacterial agents. We describe mode-of-action studies with Staphylococcus aureus of CBR-2092, a novel hybrid that combines the rifamycin SV and 4H-4-oxo-quinolizine pharmacophores. In biochemical studies, CBR-2092 exhibited rifampin-like potency as an inhibitor of RNA polymerase, was an equipotent (balanced) inhibitor of DNA gyrase and DNA topoisomerase IV, and retained activity against a prevalent quinolone-resistant variant. Macromolecular biosynthesis studies confirmed that CBR-2092 has rifampin-like effects on RNA synthesis in rifampin-susceptible strains and quinolone-like effects on DNA synthesis in rifampin-resistant strains. Studies of mutant strains that exhibited reduced susceptibility to CBR-2092 further substantiated RNA polymerase as the primary cellular target of CBR-2092, with DNA gyrase and DNA topoisomerase IV being secondary and tertiary targets, respectively, in strains exhibiting preexisting rifampin resistance. In contrast to quinolone comparator agents, no strains with altered susceptibility to CBR-2092 were found to exhibit changes consistent with altered efflux properties. The combined data indicate that CBR-2092 may have potential utility in monotherapy for the treatment of persistent S. aureus infections.

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