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Nucleic Acids Research, 2000, Vol. 28, No. 23 4604-4610
© 2000 Oxford University Press

Cloning and characterization of the Schizosaccharomyces pombe tRNA:pseudouridine synthase Pus1p

Klaus Hellmuth, Henri Grosjean1, Yuri Motorin1, Karina Deinert, Ed Hurt and George Simos*

BZH, Biochemie-Zentrum Heidelberg, Im Neuenheimer Feld 328, D-69120 Heidelberg, Germany and 1CNRS, Laboratoire d’Enzymologie et de Biochimie Structurales, 1 Avenue de la Terrasse, F-91198 Gif-sur-Yvette, France

Saccharomyces cerevisiae cells that carry deletions in both the LOS1 (a tRNA export receptor) and the PUS1 (a tRNA:pseudouridine synthase) genes exhibit a thermosensitive growth defect. A Schizosaccharomyces pombe gene, named spPUS1, was cloned from a cDNA library by complementation of this conditional lethal phenotype. The corresponding protein, spPus1p, shows sequence similarity to S.cerevisiae and murine Pus1p as well as other known members of the pseudouridine synthase family. Accordingly, recombinant spPus1p can catalyze in vitro the formation of pseudouridines at positions 27, 28, 34, 35 and 36 of yeast tRNA transcripts. The sequence and functional conservation of the Pus1p proteins in fungi and mammalian species and their notable absence from prokaryotes suggest that this family of pseudouridine synthases is required for a eukaryote-specific step of tRNA biogenesis, such as nuclear export.

* To whom correspondence should be addressed. Tel: +49 6221 546757; Fax: +49 6221 544369; Email: cg2{at}ix.urz.uni-heidelberg.de Present addresses: Klaus Hellmuth, Arimedes Biotechnology GmbH, Robert-Rössle-Straße 10, D-13125 Berlin, Germany Yuri Motorin, Maturation des ARN et Enzymologie Moléculaire, UMR 7567 CNRS-UHP, Faculté des Sciences, BP 239, 54506 Vandoeuvre-les-Nancy Cédex, France


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