Photo-affinity labelling at the peptidyl transferase centre reveals two different positions for the A- and P-sites in domain V of 23S rRNA.

ABSTRACT Oxazolidinone antibiotics inhibit bacterial protein synthesis by interacting with the large ribosomal subunit. The structure and exact location of the oxazolidinone binding site remain obscure, as does the manner in which these drugs inhibit translation. To investigate the drug-ribosome interaction, we selected Escherichia colioxazolidinone-resistant mutants, which contained a randomly mutagenized plasmid-borne rRNA operon. The same mutation, G2032 to A, was identified in the 23S rRNA genes of several independent resistant isolates. Engineering of this mutation by site-directed mutagenesis in the wild-type rRNA operon produced an oxazolidinone resistance phenotype, establishing that the G2032A substitution was the determinant of resistance. Engineered U and C substitutions at G2032, as well as a G2447-to-U mutation, also conferred resistance to oxazolidinone. All the characterized resistance mutations were clustered in the vicinity of the central loop of domain V of 23S rRNA, suggesting that this rRNA region plays a major role in the interaction of the drug with the ribosome. Although the central loop of domain V is an essential integral component of the ribosomal peptidyl transferase, oxazolidinones do not inhibit peptide bond formation, and thus these drugs presumably interfere with another activity associated with the peptidyl transferase center.

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Macrolide-Resistant and Macrolide-Sensitive Mycoplasma pneumoniae Pneumonia in Children Treated Using Early Corticosteroids

Journal of Clinical Medicine ◽

10.3390/jcm10061309 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1309

Author(s):

Hye Young Han ◽

Ki Cheol Park ◽

Eun-Ae Yang ◽

Kyung-Yil Lee

Keyword(s):

Mycoplasma Pneumoniae ◽

Serological Test ◽

Corticosteroid Treatment ◽

23S Rrna ◽

Laboratory Parameters ◽

The Mean ◽

Domain V ◽

Positive Results ◽

Mycoplasma Pneumoniae Pneumonia

We have found that early corticosteroid therapy was effective for reducing morbidity during five Korea-wide epidemics. We evaluated the clinical and laboratory parameters of 56 children who received early corticosteroid treatment for pneumonia that was caused by macrolide-resistant Mycoplasma pneumoniae (M. pneumoniae) or macrolide-sensitive M. pneumoniae between July 2019 and February 2020. All subjects had dual positive results from a PCR assay and serological test, and received corticosteroids within 24–36 h after admission. Point mutation of residues 2063, 2064, and 2067 was identified in domain V of 23S rRNA. The mean age was 6.8 years and the male:female ratio was 1.2:1 (31:25 patients). Most of the subjects had macrolide-resistant M. pneumoniae (73%), and all mutated strains had the A2063G transition. No significant differences in clinical and laboratory parameters were observed between macrolide-resistant and macrolide-sensitive M. pneumoniae groups that were treated with early dose-adjusted corticosteroids. Higher-dose steroid treatment may be needed for patients who have fever that persists for >48 h or increased biomarkers such as lactate dehydrogenase concentration at follow-up despite a usual dose of steroid therapy.

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Interaction of Arginine with the Ribosomal Peptidyl Transferase Centre

European Journal of Biochemistry ◽

10.1111/j.1432-1033.1979.tb19741.x ◽

1979 ◽

Vol 101 (2) ◽

pp. 469-473 ◽

Cited By ~ 4

Author(s):

Enrique PALACIAN ◽

David VAZQUEZ

Keyword(s):

Peptidyl Transferase ◽

Peptidyl Transferase Centre

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Structure of a conserved RNA component of the peptidyl transferase centre

Natural Structural Biology ◽

10.1038/nsb1097-775 ◽

1997 ◽

Vol 4 (10) ◽

pp. 775-778 ◽

Cited By ~ 25

Author(s):

Elisabetta Viani Puglisi ◽

Rachel Green ◽

Harry F. Noller ◽

Joseph D. Puglisi

Keyword(s):

Peptidyl Transferase ◽

Peptidyl Transferase Centre

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Protein Folding by Domain V of Escherichia coli 23S rRNA: Specificity of RNA-Protein Interactions

Journal of Bacteriology ◽

10.1128/jb.01800-07 ◽

2008 ◽

Vol 190 (9) ◽

pp. 3344-3352 ◽

Cited By ~ 36

Author(s):

Dibyendu Samanta ◽

Debashis Mukhopadhyay ◽

Saheli Chowdhury ◽

Jaydip Ghosh ◽

Saumen Pal ◽

...

Keyword(s):

Escherichia Coli ◽

Protein Folding ◽

Carbonic Anhydrase ◽

Protein Interactions ◽

23S Rrna ◽

Unfolded Proteins ◽

Bovine Carbonic Anhydrase ◽

Egg White Lysozyme ◽

General Protein ◽

Domain V

ABSTRACT The peptidyl transferase center, present in domain V of 23S rRNA of eubacteria and large rRNA of plants and animals, can act as a general protein folding modulator. Here we show that a few specific nucleotides in Escherichia coli domain V RNA bind to unfolded proteins and, as shown previously, bring the trapped proteins to a folding-competent state before releasing them. These nucleotides are the same for the proteins studied so far: bovine carbonic anhydrase, lactate dehydrogenase, malate dehydrogenase, and chicken egg white lysozyme. The amino acids that interact with these nucleotides are also found to be specific in the two cases tested: bovine carbonic anhydrase and lysozyme. They are either neutral or positively charged and are present in random coils on the surface of the crystal structure of both the proteins. In fact, two of these amino acid-nucleotide pairs are identical in the two cases. How these features might help the process of protein folding is discussed.

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Mutations in domain V of Mycoplasma pneumoniae 23S rRNA are not associated with clinical characteristics of M. pneumoniae pneumonia in children: a case control study

10.21203/rs.2.23539/v1 ◽

2020 ◽

Author(s):

Huan Deng ◽

Yifan Zhu ◽

Jiamin Zhang ◽

Qiangquan Rong ◽

Yao Quan ◽

...

Keyword(s):

Mycoplasma Pneumoniae ◽

Lymphocyte Count ◽

Clinical Characteristics ◽

Clinical Symptoms ◽

Laboratory Data ◽

Point Mutations ◽

Community Acquired Pneumonia ◽

Pulmonary Complications ◽

23S Rrna ◽

Domain V

Abstract Background Mycoplasma pneumoniae (MP) is a common agent of community-acquired pneumonia in children and young adults that can lead to refractory or persistent Mycoplasma pneumoniae pneumonia (MPP). Macrolide-resistant MP harbors point mutations in domain V of 23S ribosomal Ribonucleic Acid (rRNA) with substitutions detected at positions 2063, 2064, 2067 and 2617. This study’s purpose is to investigate the prevalence and clinical characteristics of mutations in domain V of MP 23S rRNA. Methods We sequenced the 23S rRNA domain V of MP strains collected from children with MPP. Clinical and laboratory data were also obtained, including gender, age, duration of fever, duration of fever after the start of macrolide therapy, MP-Deoxyribonucleic Acid (DNA) load at enrollment, leukocyte count, neutrophil count, and lymphocyte count, immunomodulators treatment and pulmonary complications.Results Of 276 strains, 255 (92.39 %) harbored A to G transition at the position 2063 (A2063G), and 21 (7.61 %) were not mutated. There were no significant differences in gender, age, duration of fever, duration of fever after the start of macrolide therapy, MP-DNA load at enrollment, hospitalization days, lymphocyte count and pulmonary complications when patients were stratified based on the presence or absence of domain V mutations. We also found that children with refractory MPP experienced higher MP-DNA load than the non-refractory MPP, but the prevalence of domain V mutations was comparable.Conclusions We found that clinical MP strains harbored very high mutation rate in 23S rRNA domain V, especially A2063G mutation. However, these mutations were not associated with clinical symptoms, laboratory results, pulmonary complications and development of refractory pneumonia. Instead, MP-DNA load was significantly different between refractory and non-refractory MPP.

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