| Autor: |
Reis, Filipa P., Bárria, Cátia, Gomez‐Puertas, Paulino, Gomes, Cláudio M., Arraiano, Cecília M. |
| Předmět: |
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| Zdroj: |
FEBS Letters; Feb2019, Vol. 593 Issue 3, p352-360, 9p |
| Abstrakt: |
The RNase II family of ribonucleases is ubiquitous and critical for RNA metabolism. The rnb500 allele has been widely used for over 30 years; however, the underlying genetic changes which result in RNase II thermolabile activity remain unknown. Here, we combine molecular and biophysical studies to carry out an in vivo and in vitro investigation of RNase II mutation(s) that confer the rnb500 phenotype. Our findings indicate that RNase II thermolability is due to the Cys284Tyr mutation within the RNB domain, which abolishes activity by increasing protein kinetic instability at the nonpermissive temperature. These findings have important implications for the design of temperature‐sensitive variants of other RNase II enzymes, namely those with yet unknown functions. rnb500 allele encodes a thermolabile RNase II but inherent mutations are unknown.The C284Y mutation, at a conserved position in the RNB domain, was identified.C284Y abolishes activity at the nonpermissive temperature in vivo and in vitro.The RNase II melting temperature is not affected by the C284Y mutation.C284Y increases kinetics of thermal unfolding at the nonpermissive temperature. [ABSTRACT FROM AUTHOR] |
| Databáze: |
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