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Published online 17 April 2006. doi:10.1083/jcb.200601105
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 173, Number 2, 207-218
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Article

 Stepwise RNP assembly at the site of H/ACA RNA transcription in human cells

Xavier Darzacq, Nupur Kittur, Sujayita Roy, Yaron Shav-Tal, Robert H. Singer, and U. Thomas Meier

Department of Anatomy and Structural Biology, Albert Einstein College of Medicine Bronx, NY 10461

Correspondence to Thomas Meier: meier{at}aecom.yu.edu

Mammalian H/ACA RNPs are essential for ribosome biogenesis, premessenger RNA splicing, and telomere maintenance. These RNPs consist of four core proteins and one RNA, but it is not known how they assemble. By interrogating the site of H/ACA RNA transcription, we dissected their biogenesis in single cells and delineated the role of the non-core protein NAF1 in the process. NAF1 and all of the core proteins except GAR1 are recruited to the site of transcription. NAF1 binds one of the core proteins, NAP57, and shuttles between nucleus and cytoplasm. Both proteins are essential for stable H/ACA RNA accumulation. NAF1 and GAR1 bind NAP57 competitively, suggesting a sequential interaction. Our analyses indicate that NAF1 binds NAP57 and escorts it to the nascent H/ACA RNA and that GAR1 then replaces NAF1 to yield mature H/ACA RNPs in Cajal bodies and nucleoli.

X. Darzacq, N. Kittur, and S. Roy contributed equally to this paper.

X. Darzacq's present address is Centre National de la Recherche Scientifique, École Normale Supérieure, 75230 Paris, Cedex 05, France.

Y. Shav-Tal's present address is Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan 52900, Israel.

Abbreviations used in this paper: ChIP, chromatin immunoprecipitation; CTD, COOH-terminal domain; SKL, Ser-Lys-Leu; snoRNP, small nucleolar RNP; UTR, untranslated region; SMN, survival of motor neurons; SRP, signal recognition particle.


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