Cloned (Comment) | Organism |
---|---|
recombinant overexpression of His-tagged wild-type non-acetylated and acetylated RNase II and of FLAG-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) lacking Pka or CobB | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
K501Q | site-directed mutagenesis, mutation of Lys501 results in up to 80% reduction in acetylation of RNase II | Escherichia coli |
K501R | site-directed mutagenesis, mutation of Lys501 results in up to 80% reduction in acetylation of RNase II | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | acetylation at enzyme residue Lys501 affects binding of the substrate and decreases the catalytic activity of RNase II | Escherichia coli | |
nicotinamide | NAM | Escherichia coli | |
trichostatin A | TSA | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required, no degradation of the RNA occurs in the absence of Mg2+ | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P30850 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
acetylation | residue Lys501 is acetylated in RNase II. This modification, reversibly controlled by the acetyltransferase Pka and the deacetylase CobB, a SIRT family deacetylase, affects binding of the substrate and thus decreases the catalytic activity of RNase II. Acetylation can regulate the activity of a bacterial ribonuclease. Lys501 is the major site of acetylation in this enzyme, but some other lysine residues in RNase II are also susceptible to acetylation. After enzyme inhibitor nicotinamide treatment, the acetylation level of RNase II increases 2-3fold in both wild-type and mutant strains. Inactivation of CobB increased the acetylation of wild-type RNase II | Escherichia coli |
Purification (Comment) | Organism |
---|---|
recombinant His- or FLAG-tagged wild-type non-acetylated and acetylated and mutants from Escherichia coli strain BL21(DE3) by affinity chromatography | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
RNase II | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Escherichia coli |
General Information | Comment | Organism |
---|---|---|
metabolism | role for RNase II Lys501 acetylation in modulating cell growth during stress conditions | Escherichia coli |
physiological function | RNase II is a 3' to 5' processive exoribonuclease and is the major hydrolytic enzyme in Escherichia coli accounting for about 90% of the total activity. Acetylation of residue Lys501 in RNase II, reversibly controlled by the acetyltransferase Pka and the deacetylase CobB, affects binding of the substrate and decreases the catalytic activity of RNase II. As a consequence, the steady-state level of target RNAs of RNase II may be altered in the cells. Under conditions of slowed growth, the acetylation level of RNase II is elevated and the activity of RNase II decreases, emphasizing the importance of this regulatory process | Escherichia coli |