Structural and functional analysis of Utp23, a yeast ribosome synthesis factor with degenerate PIN domain
- Jing Lu1,2,
- Mengyi Sun1,3 and
- Keqiong Ye1,2,4
- 1National Institute of Biological Sciences, Beijing, Beijing 102206, China
- 2Graduate School of Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100730, China
- 3College of Biological Sciences, China Agricultural University, Beijing 100193, China
Abstract
During synthesis of yeast ribosome, a large complex, called the 90S pre-ribosome or the small subunit processome, is assembled on the nascent precursor rRNA and mediates early processing of 18S rRNA. The Utp23 protein and snR30 H/ACA snoRNA are two conserved components of 90S pre-ribosomes. Utp23 contains a degenerate PIN nuclease domain followed by a long C-terminal tail and associates specifically with snR30. Here, we report the crystal structure of the Utp23 PIN domain at 2.5-Å resolution. The structure reveals a conserved core fold of PIN domain with degenerate active site residues, a unique CCHC Zn-finger motif, and two terminal extension elements. Functional sites of Utp23 have been examined with conservation analysis, mutagenesis, and in vivo and in vitro assays. Mutations in each of three cysteine ligands of zinc, although not the histidine ligand, were lethal or strongly inhibitory to yeast growth, indicating that the Zn-finger motif is required for Utp23 structure or function. The N-terminal helix extension harbors many highly conserved basic residues that mostly are critical for growth and in vitro RNA-binding activity of Utp23. Deletion of the C-terminal tail, which contains a short functionally important sequence motif, disrupted the interaction of Utp23 with snR30 and perturbed the pre-ribosomal association of Utp23. Our data establish a structural framework for dissecting Utp23 function in the assembly and dynamics of 90S pre-ribosomes.
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↵4 Corresponding author
E-mail yekeqiong{at}nibs.ac.cn
- Received June 23, 2013.
- Accepted September 24, 2013.
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