Previous Article | Next Article 
Molecular and Cellular Biology, December 2002, p. 8457-8466, Vol. 22, No. 24
0270-7306/02/$04.00+0 DOI: 10.1128/MCB.22.24.8457-8466.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
Immunopurified Small Nucleolar Ribonucleoprotein Particles Pseudouridylate rRNA Independently of Their Association with Phosphorylated Nopp140
Chen Wang,1 Charles C. Query,2 and U. Thomas Meier1*
Department of Anatomy and Structural Biology,1
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York 104612
Received 29 July 2002/
Returned for modification 6 September 2002/
Accepted 19 September 2002
The isomerization of up to 100 uridines to pseudouridines (
s) in eukaryotic rRNA is guided by a similar number of box H/ACA small nucleolar RNAs (snoRNAs), each forming a unique small nucleolar ribonucleoprotein particle (snoRNP) with the same four core proteins, NAP57 (also known as dyskerin or Cbf5p), GAR1, NHP2, and NOP10. Additionally, the nucleolar and Cajal body protein Nopp140 (Srp40p) associates with the snoRNPs. To understand the role of these factors in pseudouridylation, we established an in vitro assay system. Short site-specifically 32P-labeled rRNA substrates were incubated with subcellular fractions, and the conversion of uridine to
was monitored by thin-layer chromatography after digestion to single nucleotides. Immunopurified box H/ACA core particles were sufficient for the reaction. SnoRNPs associated quantitatively and reversibly with Nopp140. However, pseudouridylation activity was independent of Nopp140, consistent with a chaperoning role for this highly phosphorylated protein. Although up to 14 bp between the snoRNA and rRNA were required for the in vitro reaction, rRNA pseudouridylation and release occurred in the absence of ATP and magnesium. These data suggest that substrate release takes place without RNA helicase activity but may be aided by the snoRNP core proteins.
* Corresponding author. Mailing address: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461. Phone: (718) 430-3294. Fax: (718) 430-8996. E-mail: meier{at}aecom.yu.edu.
Molecular and Cellular Biology, December 2002, p. 8457-8466, Vol. 22, No. 24
0022-538X/02/$04.00+0 DOI: 10.1128/MCB.22.24.8457-8466.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
This article has been cited by other articles:
-
Skarie, J. M., Link, B. A.
(2008). The Primary open-angle glaucoma gene WDR36 functions in ribosomal RNA processing and interacts with the p53 stress-response pathway. Hum Mol Genet
17: 2474-2485
[Abstract]
[Full Text]
-
Montanaro, L., Trere, D., Derenzini, M.
(2008). Nucleolus, Ribosomes, and Cancer. Am. J. Pathol.
173: 301-310
[Abstract]
[Full Text]
-
Prieto, J.-L., McStay, B.
(2007). Recruitment of factors linking transcription and processing of pre-rRNA to NOR chromatin is UBF-dependent and occurs independent of transcription in human cells. Genes Dev.
21: 2041-2054
[Abstract]
[Full Text]
-
Cui, Z., DiMario, P. J.
(2007). RNAi Knockdown of Nopp140 Induces Minute-like Phenotypes in Drosophila. Mol. Biol. Cell
18: 2179-2191
[Abstract]
[Full Text]
-
Kittur, N., Zapantis, G., Aubuchon, M., Santoro, N., Bazett-Jones, D. P., Meier, U. T.
(2007). The Nucleolar Channel System of Human Endometrium Is Related to Endoplasmic Reticulum and R-Rings. Mol. Biol. Cell
18: 2296-2304
[Abstract]
[Full Text]
-
Yoon, A., Peng, G., Brandenburg, Y., Zollo, O., Xu, W., Rego, E., Ruggero, D.
(2006). Impaired control of IRES-mediated translation in X-linked dyskeratosis congenita.. Science
312: 902-906
[Abstract]
[Full Text]
-
Kelly, S., Singleton, W., Wickstead, B., Ersfeld, K., Gull, K.
(2006). Characterization and Differential Nuclear Localization of Nopp140 and a Novel Nopp140-Like Protein in Trypanosomes. Eukaryot Cell
5: 876-879
[Abstract]
[Full Text]
-
Darzacq, X., Kittur, N., Roy, S., Shav-Tal, Y., Singer, R. H., Meier, U. T.
(2006). Stepwise RNP assembly at the site of H/ACA RNA transcription in human cells. J. Cell Biol.
173: 207-218
[Abstract]
[Full Text]
-
Ballarino, M., Morlando, M., Pagano, F., Fatica, A., Bozzoni, I.
(2005). The Cotranscriptional Assembly of snoRNPs Controls the Biosynthesis of H/ACA snoRNAs in Saccharomyces cerevisiae. Mol. Cell. Biol.
25: 5396-5403
[Abstract]
[Full Text]
-
Charpentier, B., Muller, S., Branlant, C.
(2005). Reconstitution of archaeal H/ACA small ribonucleoprotein complexes active in pseudouridylation. Nucleic Acids Res
33: 3133-3144
[Abstract]
[Full Text]
-
HENRAS, A. K., CAPEYROU, R., HENRY, Y., CAIZERGUES-FERRER, M.
(2004). Cbf5p, the putative pseudouridine synthase of H/ACA-type snoRNPs, can form a complex with Gar1p and Nop10p in absence of Nhp2p and box H/ACA snoRNAs. RNA
10: 1704-1712
[Abstract]
[Full Text]
-
Mochizuki, Y., He, J., Kulkarni, S., Bessler, M., Mason, P. J.
(2004). Mouse dyskerin mutations affect accumulation of telomerase RNA and small nucleolar RNA, telomerase activity, and ribosomal RNA processing. Proc. Natl. Acad. Sci. USA
101: 10756-10761
[Abstract]
[Full Text]
-
Henning, D., So, R. B., Jin, R., Lau, L. F., Valdez, B. C.
(2003). Silencing of RNA Helicase II/Gu{alpha} Inhibits Mammalian Ribosomal RNA Production. J. Biol. Chem.
278: 52307-52314
[Abstract]
[Full Text]
-
Yang, Y., Meier, U. T.
(2003). Genetic Interaction between a Chaperone of Small Nucleolar Ribonucleoprotein Particles and Cytosolic Serine Hydroxymethyltransferase. J. Biol. Chem.
278: 23553-23560
[Abstract]
[Full Text]
Copyright © 2002 by the American Society for Microbiology. All rights reserved.