3U28

Crystal structure of a Cbf5-Nop10-Gar1 complex from Saccharomyces cerevisiae

Summary for 3U28

• Entry DOI: 10.2210/pdb3u28/pdb
• Descriptor: H/ACA ribonucleoprotein complex subunit 4, H/ACA ribonucleoprotein complex subunit 3, H/ACA ribonucleoprotein complex subunit 1, ... (4 entities in total)
• Functional Keywords: pseudouridine synthase, pseudouridylation, h/aca rna, nucleolus, isomerase-protein binding complex, isomerase/protein binding
• Biological source: Saccharomyces cerevisiae (yeast); More
• Cellular location: Nucleus, nucleolus: P33322 Q6Q547 P28007
• Total number of polymer chains: 3
• Total formula weight: 64249.97

Authors

Ye, K.,Li, S. (deposition date: 2011-10-02, release date: 2011-12-14, Last modification date: 2023-11-01)

Primary citation

Li, S.,Duan, J.,Li, D.,Yang, B.,Dong, M.,Ye, K.
Reconstitution and structural analysis of the yeast box H/ACA RNA-guided pseudouridine synthase
Genes Dev., 25:2409-2421, 2011

PubMed Abstract: Box H/ACA ribonucleoprotein particles (RNPs) mediate pseudouridine synthesis, ribosome formation, and telomere maintenance. The structure of eukaryotic H/ACA RNPs remains poorly understood. We reconstituted functional Saccharomyces cerevisiae H/ACA RNPs with recombinant proteins Cbf5, Nop10, Gar1, and Nhp2 and a two-hairpin H/ACA RNA; determined the crystal structure of a Cbf5, Nop10, and Gar1 ternary complex at 1.9 Å resolution; and analyzed the structure-function relationship of the yeast complex. Although eukaryotic H/ACA RNAs have a conserved two-hairpin structure, isolated single-hairpin RNAs are also active in guiding pseudouridylation. Nhp2, unlike its archaeal counterpart, is largely dispensable for the activity, reflecting a functional adaptation of eukaryotic H/ACA RNPs to the variable RNA structure that Nhp2 binds. The N-terminal extension of Cbf5, a hot spot for dyskeratosis congenita mutation, forms an extra structural layer on the PUA domain. Gar1 is distinguished from the assembly factor Naf1 by containing a C-terminal extension that controls substrate turnover and the Gar1-Naf1 exchange during H/ACA RNP maturation. Our results reveal significant novel features of eukaryotic H/ACA RNPs.

PubMed: 22085967
DOI: 10.1101/gad.175299.111

Experimental method

X-RAY DIFFRACTION (1.9 Å)