Purification of Terminal Uridylyltransferase from the Crithidia Kinetoplast-Mitochondrion (RESEARCH FACILITY OF NUCLEIC ACIDS)

Abstract

Terminal uridylyltransferase is a candidate responsible for a post-transcriptional RNA editing process of mitochondrial transcripts in kinetoplastid protozoans. The activity was solubilized by detergent lysis of mitochondria of Crithidia fasciculata isolated by the Renografin density gradient floatation centrifugation, and the enzyme was partially purified using chromatography on phosphocellulose, DEAE-cellulose and Sephadex G200 columns. The major activity of the purified enzyme was to add more than 30 uridylates to the 3’-OH end of guide RNA molecules, required magnesium cations and acted in a UTP dependent manner.