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Originally published In Press as doi:10.1074/jbc.M203399200 on June 4, 2002

J. Biol. Chem., Vol. 277, Issue 32, 28780-28786, August 9, 2002
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Rpf2p, an Evolutionarily Conserved Protein, Interacts with Ribosomal Protein L11 and Is Essential for the Processing of 27 SB Pre-rRNA to 25 S rRNA and the 60 S Ribosomal Subunit Assembly in Saccharomyces cerevisiae*

Daisuke MoritaDagger , Keita MiyoshiDagger §, Yasushi Matsui||, Akio Toh-e||, Hidenori ShinkawaDagger , Tokichi MiyakawaDagger , and Keiko MizutaDagger §**

From the Dagger  Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter and the § Department of Bioresource Science and Technology, Graduate School of Biosphere Sciences, Hiroshima University, Kagamiyama 1-4-4, Higashi-Hiroshima 739-8528, Japan and the || Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan

Saccharomyces cerevisiae Rrs1p is a nuclear protein that is essential for the maturation of 25 S rRNA and the 60 S ribosomal subunit assembly. In two-hybrid screening, using RRS1 as bait, we have cloned YKR081c/RPF2. Rpf2p is essential for growth and is mainly localized in the nucleolus. The amino acid sequence of Rpf2p is highly conserved in eukaryotes from yeast to human. Similar to Rrs1p, Rpf2p shows physical interaction with ribosomal protein L11 and appears to associate with preribosomal subunits fairly tightly. Northern, methionine pulse-chase, and sucrose density gradient ultracentrifugation analyses reveal that the depletion of Rpf2p results in a delayed processing of pre-rRNA, a decrease of mature 25 S rRNA, and a shortage of 60 S subunits. An analysis of processing intermediates by primer extension shows that the Rpf2p depletion leads to an accumulation of 27 SB pre-rRNA, suggesting that Rpf2p is required for the processing of 27 SB into 25 S rRNA.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Recipient of a Japan Society for the Promotion of Science predoctoral research fellowship.

** To whom correspondence should be addressed. Tel.: 81-824-24-7926; Fax: 81-824-24-7926; E-mail: kmizuta@hiroshima-u.ac.jp.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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