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Molecular and Cellular Biology, July 2004, p. 5797-5807, Vol. 24, No. 13
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.13.5797-5807.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Human Box H/ACA Pseudouridylation Guide RNA Machinery{dagger}

Arnold M. Kiss,1,2 Beáta E. Jády,2,3 Edouard Bertrand,3 and Tamás Kiss1,2*

Laboratoire de Biologie Moléculaire Eucaryote du CNRS, UMR5099, IFR109 CNRS, 31062 Toulouse Cedex 4,1 Institut de Génétique Moléculaire, 34000 Montpellier, France,3 Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary2

Received 26 January 2004/ Returned for modification 23 March 2004/ Accepted 1 April 2004

Pseudouridine, the most abundant modified nucleoside in RNA, is synthesized by posttranscriptional isomerization of uridines. In eukaryotic RNAs, site-specific synthesis of pseudouridines is directed primarily by box H/ACA guide RNAs. In this study, we have identified 61 novel putative pseudouridylation guide RNAs by construction and characterization of a cDNA library of human box H/ACA RNAs. The majority of the new box H/ACA RNAs are predicted to direct pseudouridine synthesis in rRNAs and spliceosomal small nuclear RNAs. We can attribute RNA-directed modification to 79 of the 97 pseudouridylation sites present in the human 18S, 5.8S, and 28S rRNAs and to 11 of the 21 pseudouridines reported for the U1, U2, U4, U5, and U6 spliceosomal RNAs. We have also identified 12 novel box H/ACA RNAs which lack apparent target pseudouridines in rRNAs and small nuclear RNAs. These putative guide RNAs likely function in the pseudouridylation of some other types of cellular RNAs, suggesting that RNA-guided pseudouridylation is more general than assumed before. The genomic organization of the new box H/ACA RNA genes indicates that in human cells, all box H/ACA pseudouridylation guide RNAs are processed from introns of pre-mRNA transcripts which either encode a protein product or lack protein-coding capacity.

* Corresponding author. Mailing address: Laboratoire de Biologie Moléculaire Eucaryote du CNRS, 118 Rt. de Narbonne, 31062 Toulouse Cedex, France. Phone: 33 5 61 33 59 91. Fax: 33 5 61 33 58 86. E-mail: tamas{at}ibcg.biotoul.fr.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, July 2004, p. 5797-5807, Vol. 24, No. 13
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.13.5797-5807.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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