Assays of the helicase, ATPase, and exoribonuclease activities of the yeast mitochondrial degradosome.
The mitochondrial degradosome (mtEXO) is the main enzymatic complex in RNA degradation, processing, and surveillance in Saccharomyces cerevisiae mitochondria. It consists of two nuclear-encoded subunits: the ATP-dependent RNA helicase Suv3p and the 3' to 5' exoribonuclease Dss1p. The two subunits depend on each other for their activity; the complex can therefore be considered as a model system for the cooperation of RNA helicases and exoribonucleases in RNA degradation. All the three activities of the complex (helicase, ATPase, and exoribonuclease) can be studied in vitro using recombinant proteins and protocols presented in this chapter.
Malecki
Michal
M
Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw, Pawinskiego 5A, 02-106, Warsaw, Poland.
Stepien
Piotr P
PP
Golik
Pawel
P
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D001681:Biological Assay / Q000379:methods*; D004722:Endoribonucleases / Q000737:chemistry / Q000378:metabolism*; D005095:Exoribonucleases / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology*; D009097:Multienzyme Complexes / Q000737:chemistry / Q000378:metabolism*; D011117:Polyribonucleotide Nucleotidyltransferase / Q000737:chemistry / Q000378:metabolism*; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D020365:RNA Helicases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
D009097:Multienzyme Complexes; D021122:Protein Subunits; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C111244:degradosome; D011117:Polyribonucleotide Nucleotidyltransferase; D004722:Endoribonucleases; D005095:Exoribonucleases; D000251:Adenosine Triphosphatases; D020365:RNA Helicases