Cloned (Comment) | Organism |
---|---|
wild-type and RNase II mutants cloned into plasmid pFCT6.9 and expressed in Escherichia coli BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D201N | significant loss of activity in degradation of poly(A) (0.2% of that of the wild-type enzyme). Generates a 10-11-nt fragment as a major degradation product, although longer reaction times result in the usual 4-nt fragment as a secondary product | Escherichia coli |
D207N | still retains 12% activity | Escherichia coli |
D210N | significant loss of activity in degradation of poly(A) (0.3% of that of the wild-type enzyme). Generates a 10-11-nt fragment as a major degradation product, although longer reaction times result in the usual 4-nt fragment as a secondary product | Escherichia coli |
F358A | the protein is 2fold more active than the wild-type | Escherichia coli |
Y253A | 26% of the activity of the enzyme persists, significantly impairs RNA binding | Escherichia coli |
Y253A/F358A | 12% of the activity of the enzyme persists, whereas RNA binding affinity is not significantly affected | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | Asp209, but not Asp207, contacting Mg2+ in addition to Asp201, Asp210, and the phosphate group of nt 13 | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P30850 | - |
- |
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
wild-type and RNase II mutants purified by affinity chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Tyr253 and Phe358 are not essential for catalysis by RNase II. Tyr253 seems to be a critical residue in setting the smallest product generated by RNase II, thus being important for the stabilization of the 3'-end of the RNA molecule. Tyr253 is highly conserved and equivalent residues are present in many RNase II family members. Phe358 can be preventing a faster degradation of the RNA by stalling its translocation, probably due to the stacking of its aromatic ring between the bases of contiguous nucleotides. Asp201, Asp207, Asp209, and Asp210 are located in the RNase II active site. These residues are not equivalent and their functions in RNA metabolism are distinct, whereby Asp209 is the only residue essential for RNase II activity | Escherichia coli | ? | - |
? | |
oligoribonucleotide + H2O | final end product of RNase II is 4-nt, whereas for RNase R it is a 2-nt fragment | Escherichia coli | ? | - |
? | |
oligoribonucleotide + H2O | final end product of Rrp44 is 4-nt | Homo sapiens | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | human Rrp44 proteins shows an overall structure similar to that of Escherichia coli RNase II | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
Dis3 | - |
Homo sapiens |
ribonuclease II | - |
Homo sapiens |
ribonuclease II | - |
Escherichia coli |
RNase II | - |
Homo sapiens |
RNase II | - |
Escherichia coli |
RNase R | - |
Escherichia coli |
Rrp44 | - |
Homo sapiens |