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Structural characterization of the ribosomal P1A-P2B protein dimer by small-angle X-ray scattering and NMR spectroscopy

Grela, Przemysłw; Helgstrand, Magnus LU ; Krokowski, Dawid; Boguszewska, Aleksandra; Svergun, Dmitri; Liljas, Anders LU ; Bernado, Pau; Grankowski, Nikodem; Akke, Mikael LU and Tchorzewski, Marek (2007) In Biochemistry 46(7). p.1988-1998
Abstract
The five ribosomal P-proteins, denoted P0-(P1-P2)(2), constitute the stalk structure of the large subunit of eukaryotic ribosomes. In the yeast Saccharomyces cerevisiae, the group of P1 and P2 proteins is differentiated into subgroups that form two separate P1A-P2B and P1B-P2A heterodimers on the stalk. So far, structural studies on the P-proteins have not yielded any satisfactory information using either X-ray crystallography or NMR spectroscopy, and the structures of the ribosomal stalk and its individual constituents remain obscure. Here we outline a first, coarse-grained view of the P1A-P2B solution structure obtained by a combination of small-angle X-ray scattering and heteronuclear NMR spectroscopy. The complex has an elongated shape... (More)
The five ribosomal P-proteins, denoted P0-(P1-P2)(2), constitute the stalk structure of the large subunit of eukaryotic ribosomes. In the yeast Saccharomyces cerevisiae, the group of P1 and P2 proteins is differentiated into subgroups that form two separate P1A-P2B and P1B-P2A heterodimers on the stalk. So far, structural studies on the P-proteins have not yielded any satisfactory information using either X-ray crystallography or NMR spectroscopy, and the structures of the ribosomal stalk and its individual constituents remain obscure. Here we outline a first, coarse-grained view of the P1A-P2B solution structure obtained by a combination of small-angle X-ray scattering and heteronuclear NMR spectroscopy. The complex has an elongated shape with a length of 10 nm and a cross section of similar to 2.5 nm. N-15 NMR relaxation measurements establish that roughly 30% of the residues are present in highly flexible segments, which belong primarily to the linker region and the C-terminal part of the polypeptide chain. Secondary structure predictions and NMR chemical shift analysis, together with previous results from CD spectroscopy, indicate that the structured regions involve alpha-helices. NMR relaxation data further suggest that several helices are arranged in a nearly parallel or antiparallel topology. These results provide the first structural comparison between eukaryotic P1 and P2 proteins and the prokaryotic L12 counterpart, revealing considerable differences in their overall shapes, despite similar functional roles and similar oligomeric arrangements. These results present for the first time a view of the structure of the eukaryotic stalk constituents, which is the only domain of the eukaryotic ribosome that has escaped successful structural characterization. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biochemistry
volume
46
issue
7
pages
1988 - 1998
publisher
The American Chemical Society
external identifiers
  • wos:000244161900029
  • scopus:33847046017
ISSN
0006-2960
DOI
10.1021/bi0616450
language
English
LU publication?
yes
id
d2e9d13f-d7a0-44e6-bbee-60be12f308c6 (old id 572454)
date added to LUP
2007-10-31 11:45:10
date last changed
2017-08-27 03:50:48
@article{d2e9d13f-d7a0-44e6-bbee-60be12f308c6,
  abstract     = {The five ribosomal P-proteins, denoted P0-(P1-P2)(2), constitute the stalk structure of the large subunit of eukaryotic ribosomes. In the yeast Saccharomyces cerevisiae, the group of P1 and P2 proteins is differentiated into subgroups that form two separate P1A-P2B and P1B-P2A heterodimers on the stalk. So far, structural studies on the P-proteins have not yielded any satisfactory information using either X-ray crystallography or NMR spectroscopy, and the structures of the ribosomal stalk and its individual constituents remain obscure. Here we outline a first, coarse-grained view of the P1A-P2B solution structure obtained by a combination of small-angle X-ray scattering and heteronuclear NMR spectroscopy. The complex has an elongated shape with a length of 10 nm and a cross section of similar to 2.5 nm. N-15 NMR relaxation measurements establish that roughly 30% of the residues are present in highly flexible segments, which belong primarily to the linker region and the C-terminal part of the polypeptide chain. Secondary structure predictions and NMR chemical shift analysis, together with previous results from CD spectroscopy, indicate that the structured regions involve alpha-helices. NMR relaxation data further suggest that several helices are arranged in a nearly parallel or antiparallel topology. These results provide the first structural comparison between eukaryotic P1 and P2 proteins and the prokaryotic L12 counterpart, revealing considerable differences in their overall shapes, despite similar functional roles and similar oligomeric arrangements. These results present for the first time a view of the structure of the eukaryotic stalk constituents, which is the only domain of the eukaryotic ribosome that has escaped successful structural characterization.},
  author       = {Grela, Przemysłw and Helgstrand, Magnus and Krokowski, Dawid and Boguszewska, Aleksandra and Svergun, Dmitri and Liljas, Anders and Bernado, Pau and Grankowski, Nikodem and Akke, Mikael and Tchorzewski, Marek},
  issn         = {0006-2960},
  language     = {eng},
  number       = {7},
  pages        = {1988--1998},
  publisher    = {The American Chemical Society},
  series       = {Biochemistry},
  title        = {Structural characterization of the ribosomal P1A-P2B protein dimer by small-angle X-ray scattering and NMR spectroscopy},
  url          = {http://dx.doi.org/10.1021/bi0616450},
  volume       = {46},
  year         = {2007},
}