Abstract：
:A C-->U mutation (rdn5) in the conserved sarcin/ricin domain of yeast 25S rRNA has been shown to cause translational suppression and paromomycin resistance. It also separates the killing from the misreading effect of this antibiotic. We confirm these findings and provide in vitro evidence that rdn5 causes a 3-fold increase in translational errors and resistance to paromomycin. The role of this 25S rRNA domain in ribosome's decoding function was further demonstrated when 60S subunits from rdn5 cells were combined with 40S subunits from cells carrying an error-prone mutation in the eukaryotic accuracy center ribosomal protein S23, an homologue of Escherichia coli S12. These hybrids exhibited an error frequency similar to that of rdn5 alone, despite the error-prone mutation in S23. This was accompanied by extreme resistance to paromomycin, unlike the effects of the individual mutations. Furthermore, rdn5 lowers peptidyltransferase activity measured as a second-order rate constant (kcat/K(s)) corresponding to the rate of peptide bond formation. This mutation was also found to affect translocation. Elongation factor 2 (EF2)-dependent translocation of Ac-Phe-tRNA from the A- to P-site was achieved at an EF2 concentration 3.5 times lower than in wild type. In conclusion, the sarcin/ricin domain of 25S rRNA influences decoding, peptide bond formation and translocation.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Panopoulos P,Dresios J,Synetos Ddoi
10.1093/nar/gkh860keywords:
subject
Has Abstractpub_date
2004-10-11 00:00:00pages
5398-408issue
18eissn
0305-1048issn
1362-4962pii
32/18/5398journal_volume
32pub_type
杂志文章abstract：:Cloned DNA from the EcoRI J fragment of EBV has been used as template for in vitro transcription experiments using cell-free extracts prepared from HeLa or KB cells. Two EBV specific RNAs each about 175 bases in length were synthesised and nuclease 51 mapping experiments determined that these in vitro products corresp...
journal_title：Nucleic acids research
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doi：10.1093/nar/10.11.3407
更新日期：1982-06-11 00:00:00
abstract：:The complete nucleotide sequence of the Escherichia coli recB gene which encodes a subunit of the ATP-dependent DNase, Exonuclease V, has been determined. The proposed coding region for the RecB protein is 3543 nucleotides long and would encode a polypeptide of 1180 amino acids with a calculated molecular weight of 13...
journal_title：Nucleic acids research
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更新日期：1986-11-11 00:00:00
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journal_title：Nucleic acids research
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pub_type： 杂志文章
doi：10.1093/nar/22.3.427
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journal_title：Nucleic acids research
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更新日期：2009-12-01 00:00:00
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更新日期：1988-10-11 00:00:00
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journal_title：Nucleic acids research
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更新日期：2018-09-28 00:00:00
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journal_title：Nucleic acids research
pub_type： 杂志文章
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更新日期：2013-12-01 00:00:00
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journal_title：Nucleic acids research
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更新日期：2017-10-13 00:00:00
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journal_title：Nucleic acids research
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更新日期：2005-07-01 00:00:00
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journal_title：Nucleic acids research
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journal_title：Nucleic acids research
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更新日期：1990-04-11 00:00:00
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