Synthesis and Assembly of the Box C+D Small Nucleolar RNPs

doi:10.1128/MCB.20.8.2650-2659.2000

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Molecular and Cellular Biology, April 2000, p. 2650-2659, Vol. 20, No. 8
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Synthesis and Assembly of the Box C+D Small Nucleolar RNPs

Denis L. J. Lafontaine^* and David Tollervey

ICMB, The University of Edinburgh, Edinburgh EH9 3JR, Scotland

Received 9 September 1999/Returned for modification 25 October 1999/Accepted 10 January 2000

Two core small nucleolar RNP (snoRNP) proteins, Nop1p (fibrillarin in vertebrates) and Nop58p (also known as Nop5p) have previouslybeen reported to be specifically associated with the box C+D classof small nucleolar RNAs (snoRNAs). Here we report that Nop56p,a protein related in sequence to Nop58p, is a bona fide box C+DsnoRNP component; all tested box C+D snoRNAs were coprecipitatedwith protein A-tagged Nop56p. Analysis of in vivo snoRNP assemblyindicated that Nop56p was stably associated with the snoRNAs onlyin the presence of Nop1p. In contrast, Nop58p and Nop1p associateindependently with the snoRNAs. Genetic depletion of Nop56p resultedin inhibition of early pre-rRNA processing events at sites A₀,A₁, and A₂ and mild depletion of 18S rRNA. However, Nop56p depletiondid not lead to codepletion of the box C+D snoRNAs. This is incontrast to Nop58p, which was required for the accumulation ofall tested box C+D snoRNAs. Unexpectedly, we found that Nop1pwas specifically required for the synthesis and accumulation ofbox C+D snoRNAs processed from pre-mRNA introns and polycistronictranscripts.

^* Corresponding author. Mailing address: Institute of Cell and Molecular Biology, Swann Building, King's Buildings, The Universityof Edinburgh, Mayfield Rd., EH9 3JR Edinburgh, Scotland. Phone:44 131 650 7093. Fax: 44 131 650 7040 or 8650. E-mail: denis.lafontaine{at}ed.ac.uk.

Molecular and Cellular Biology, April 2000, p. 2650-2659, Vol. 20, No. 8
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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