The human RNA 3'-terminal phosphate cyclase is a member of a new family of proteins conserved in Eucarya, Bacteria and Archaea.
RNA 3'-terminal phosphate cyclase catalyses the ATP-dependent conversion of the 3'-phosphate to a 2',3'-cyclic phosphodiester at the end of RNA. The physiological function of the cyclase is not known, but the enzyme could be involved in the maintenance of cyclic ends in tRNA splicing intermediates or in the cyclization of the 3' end of U6 snRNA. In this work, we describe cloning of the human cyclase cDNA. The purified bacterially overexpressed protein underwent adenylylation in the presence of [alpha-32P]ATP and catalysed cyclization of the 3'-terminal phosphate in different RNA substrates, consistent with previous findings. Comparison of oligoribonucleotides and oligodeoxyribonucleotides of identical sequence demonstrated that the latter are approximately 500-fold poorer substrates for the enzyme. In Northern analysis, the cyclase was expressed in all analysed mammalian tissues and cell lines. Indirect immunofluorescence, performed with different transfected mammalian cell lines, showed that this protein is nuclear, with a diffuse nucleoplasmic localization. The sequence of the human cyclase has no apparent motifs in common with any proteins of known function. However, inspection of the databases identified proteins showing strong similarity to the enzyme, originating from as evolutionarily distant organisms as yeast, plants, the bacterium Escherichia coli and the archaeon Methanococcus jannaschii. The overexpressed E. coli protein has cyclase activity similar to that of the human enzyme. The conservation of the RNA 3'-terminal phosphate cyclase among Eucarya, Bacteria and Archaea argues that the enzyme performs an important function in RNA metabolism.
Genschik
P
P
Friedrich Miescher-Institut, Basel, Switzerland.
Billy
E
E
Swianiewicz
M
M
Filipowicz
W
W
D000595:Amino Acid Sequence; D000818:Animals; D001105:Archaea / Q000201:enzymology / Q000235:genetics; D001419:Bacteria / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000737:chemistry; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005057:Eukaryotic Cells; D005455:Fluorescent Antibody Technique; D006801:Humans; D008025:Ligases / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D011387:Prokaryotic Cells; D012313:RNA / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013379:Substrate Specificity; D014018:Tissue Distribution
D009687:Nuclear Proteins; D011994:Recombinant Proteins; D012313:RNA; D008025:Ligases; C037451:RNA 3'-terminal phosphate cyclase