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Molecular and Cellular Biology, April 2006, p. 2540-2549, Vol. 26, No. 7
0270-7306/06/$08.00+0     doi:10.1128/MCB.26.7.2540-2549.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Cotranscriptional Recognition of Human Intronic Box H/ACA snoRNAs Occurs in a Splicing-Independent Manner

Patricia Richard,1 Arnold M. Kiss,1,2 Xavier Darzacq,1 and Tamás Kiss1,2*

Laboratoire de Biologie Moléculaire Eucaryote du CNRS, UMR5099, IFR109, 118 route de Narbonne, 31062 Toulouse, France,1 Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary2

Received 15 September 2005/ Returned for modification 21 October 2005/ Accepted 13 January 2006

Processing from pre-mRNA introns is a widespread mechanism to generate human box C/D and H/ACA snoRNAs. Recent studies revealed that an optimal position relative to the 3' splice site is important for efficient processing of most box C/D snoRNAs and that assembly of box C/D snoRNPs is stimulated by splicing factors likely bound to the branch point region. Here we have investigated the processing of another major class of human intron-encoded RNAs, the box H/ACA snoRNAs. Analysis of 80 H/ACA RNA genes revealed that human H/ACA RNAs possess no preferential localization close to the 3' or 5' splice site. In vivo processing experiments confirmed that H/ACA intronic snoRNAs are processed in a position-independent manner, indicating that there is no synergy between H/ACA RNA processing and splicing. We also showed that recognition of intronic H/ACA snoRNAs and assembly of pre-snoRNPs is an early event that occurs during transcription elongation parallel with pre-mRNA splice site selection. Finally, we found that efficient processing and correct nucleolar localization of the human U64 H/ACA snoRNA requires RNA polymerase II-mediated synthesis of the U64 precursor. This suggests that polymerase II-associated factors direct the efficient assembly and determine the correct subnuclear trafficking of human H/ACA snoRNPs.

* Corresponding author. Mailing address: Laboratoire de Biologie Moléculaire Eucaryote du CNRS, UMR5099, IFR109, 118 route de Narbonne, 31062 Toulouse, France. Phone: (33) 5-61-33-59-07. Fax: (33) 5-61-33-58-86. E-mail: tamas{at}ibcg.biotoul.fr.

Molecular and Cellular Biology, April 2006, p. 2540-2549, Vol. 26, No. 7
0022-538X/06/$08.00+0     doi:10.1128/MCB.26.7.2540-2549.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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