Cloned (Comment) | Organism |
---|---|
full-length Rrp44 (residues 1-1001) and truncated constructs of Rrp44 expressed from a pETM11 vector in Escherichia coli | Saccharomyces cerevisiae |
Crystallization (Comment) | Organism |
---|---|
Rrp44 in complex with single-stranded RNA, to 2.3 A resolution. Structure of Rrp44 displays CSD1, CSD2, RNB, and S1 domains. The two N-terminal cold shock domains (CSD1, residues 271-399; CSD2, residues 400-475) and the C-terminal S1 domain (residues 911-998) display characteristic OB folds, with five antiparallel beta strands organized in a beta barrel structure. CSD1 is fused to an N-terminal alpha helix (residues 261-268) and the S1 domain has an insertion of three beta strands (between beta3 and beta4) as compared to a standard OB fold. The RNB domain is centered around a core and is surrounded by several alpha helices | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
A815F | degrades RNA duplexes with 7 or 14 nucleotides of ssRNA overhang significantly slower (about 4fold) than the wild-type enzyme | Saccharomyces cerevisiae |
A815W | degrades RNA duplexes with 7 or 14 nucleotides of ssRNA overhang significantly slower (about 3fold) than the wild-type enzyme | Saccharomyces cerevisiae |
D551N | abolishes the exonucleolytic activity | Saccharomyces cerevisiae |
additional information | Rrp44 242-1001 (Rrp44DELTAN) lacks the predicted N-terminal PIN domain, shows no detectable difference in activity toward ssRNA substrates as compared to recombinant Rrp44 | Saccharomyces cerevisiae |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
additional information | exosome, i.e. a macromolecule complex involved in RNA degradation | Saccharomyces cerevisiae | - |
- |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | Q08162 | - |
- |
Purification (Comment) | Organism |
---|---|
full-length protein and mutants purified by affinity Ni-NTA chromatography, followed by ion exchange chromatography and gel filtration. 242-1001 construct purified to homogeneity | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dsRNA + H2O | Rrp44 is very efficient in degrading a duplex with a 30 overhang of 14 nucleotides and is inactive with overhangs as short as 2 or 3 nucleotides | Saccharomyces cerevisiae | ? | - |
? | |
ssRNA + H2O | 50fold higher affinity for ssRNA than for a corresponding ssDNA oligonucleotide | Saccharomyces cerevisiae | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Rrp44 | - |
Saccharomyces cerevisiae |