E. coli RNase I exhibits a strong Ca2+-dependent inherent double-stranded RNase activity
© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research..
Since its initial characterization, Escherichia coli RNase I has been described as a single-strand specific RNA endonuclease that cleaves its substrate in a largely sequence independent manner. Here, we describe a strong calcium (Ca2+)-dependent activity of RNase I on double-stranded RNA (dsRNA), and a Ca2+-dependent novel hybridase activity, digesting the RNA strand in a DNA:RNA hybrid. Surprisingly, Ca2+ does not affect the activity of RNase I on single stranded RNA (ssRNA), suggesting a specific role for Ca2+ in the modulation of RNase I activity. Mutation of a previously overlooked Ca2+ binding site on RNase I resulted in a gain-of-function enzyme that is highly active on dsRNA and could no longer be stimulated by the metal. In summary, our data imply that native RNase I contains a bound Ca2+, allowing it to target both single- and double-stranded RNAs, thus having a broader substrate specificity than originally proposed for this traditional enzyme. In addition, the finding that the dsRNase activity, and not the ssRNase activity, is associated with the Ca2+-dependency of RNase I may be useful as a tool in applied molecular biology.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:49 |
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| Enthalten in: |
Nucleic acids research - 49(2021), 9 vom: 21. Mai, Seite 5265-5277 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Grünberg, Sebastian [VerfasserIn] |
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| Links: |
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| Themen: |
63231-63-0 |
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| Anmerkungen: |
Date Completed 06.07.2021 Date Revised 06.07.2021 published: Print Citation Status MEDLINE |
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| doi: |
10.1093/nar/gkab284 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM324400055 |
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| 245 | 1 | 0 | |a E. coli RNase I exhibits a strong Ca2+-dependent inherent double-stranded RNase activity |
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| 520 | |a © The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research. | ||
| 520 | |a Since its initial characterization, Escherichia coli RNase I has been described as a single-strand specific RNA endonuclease that cleaves its substrate in a largely sequence independent manner. Here, we describe a strong calcium (Ca2+)-dependent activity of RNase I on double-stranded RNA (dsRNA), and a Ca2+-dependent novel hybridase activity, digesting the RNA strand in a DNA:RNA hybrid. Surprisingly, Ca2+ does not affect the activity of RNase I on single stranded RNA (ssRNA), suggesting a specific role for Ca2+ in the modulation of RNase I activity. Mutation of a previously overlooked Ca2+ binding site on RNase I resulted in a gain-of-function enzyme that is highly active on dsRNA and could no longer be stimulated by the metal. In summary, our data imply that native RNase I contains a bound Ca2+, allowing it to target both single- and double-stranded RNAs, thus having a broader substrate specificity than originally proposed for this traditional enzyme. In addition, the finding that the dsRNase activity, and not the ssRNase activity, is associated with the Ca2+-dependency of RNase I may be useful as a tool in applied molecular biology | ||
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| 700 | 1 | |a Coxam, Baptiste |e verfasserin |4 aut | |
| 700 | 1 | |a Chen, Tien-Hao |e verfasserin |4 aut | |
| 700 | 1 | |a Dai, Nan |e verfasserin |4 aut | |
| 700 | 1 | |a Saleh, Lana |e verfasserin |4 aut | |
| 700 | 1 | |a Corrêa, Ivan R |e verfasserin |4 aut | |
| 700 | 1 | |a Nichols, Nicole M |e verfasserin |4 aut | |
| 700 | 1 | |a Yigit, Erbay |e verfasserin |4 aut | |
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