Insights into How RNase R Degrades Structured RNA: Analysis of the Nuclease Domain

Helen A. Vincent, Murray P Deutscher

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

RNase R readily degrades highly structured RNA, whereas its paralogue, RNase II, is unable to do so. Furthermore, the nuclease domain of RNase R, devoid of all canonical RNA-binding domains, is sufficient for this activity. RNase R also binds RNA more tightly within its catalytic channel than does RNase II, which is thought to be important for its unique catalytic properties. To investigate this idea further, certain residues within the nuclease domain channel of RNase R were changed to those found in RNase II. Among the many examined, we identified one amino acid residue, R572, that has a significant role in the properties of RNase R. Conversion of this residue to lysine, as found in RNase II, results in weaker substrate binding within the nuclease domain channel, longer limit products, increased activity against a variety of substrates and a faster substrate on-rate. Most importantly, the mutant encounters difficulty in degrading structured RNA, pausing within a double-stranded region. Additional studies show that degradation of structured substrates is dependent upon temperature, suggesting a role for thermal breathing in the mechanism of action of RNase R. On the basis of these data, we propose a model in which tight binding within the nuclease domain allows RNase R to capitalize on the natural thermal breathing of an RNA duplex to degrade structured RNAs.

Original languageEnglish
Pages (from-to)570-583
Number of pages14
JournalJournal of Molecular Biology
Volume387
Issue number3
DOIs
StatePublished - Apr 3 2009

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Ribonucleases
RNA
Respiration
Hot Temperature
ribonuclease R
Lysine
Amino Acids
Temperature
exoribonuclease II

Keywords

  • Escherichia coli ribonuclease
  • RNA binding
  • RNA degradation
  • RNase II
  • RNase R

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Insights into How RNase R Degrades Structured RNA : Analysis of the Nuclease Domain. / Vincent, Helen A.; Deutscher, Murray P.

In: Journal of Molecular Biology, Vol. 387, No. 3, 03.04.2009, p. 570-583.

Research output: Contribution to journalArticle

Vincent, Helen A. ; Deutscher, Murray P. / Insights into How RNase R Degrades Structured RNA : Analysis of the Nuclease Domain. In: Journal of Molecular Biology. 2009 ; Vol. 387, No. 3. pp. 570-583.
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