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Title: Structural and functional analysis of the human spliceosomal DEAD-box helicase Prp28

The crystal structure of the helicase domain of the human spliceosomal DEAD-box protein Prp28 was solved by SAD. The binding of ADP and ATP by Prp28 was studied biochemically and analysed with regard to the crystal structure. The DEAD-box protein Prp28 is essential for pre-mRNA splicing as it plays a key role in the formation of an active spliceosome. Prp28 participates in the release of the U1 snRNP from the 5′-splice site during association of the U5·U4/U6 tri-snRNP, which is a crucial step in the transition from a pre-catalytic spliceosome to an activated spliceosome. Here, it is demonstrated that the purified helicase domain of human Prp28 (hPrp28ΔN) binds ADP, whereas binding of ATP and ATPase activity could not be detected. ATP binding could not be observed for purified full-length hPrp28 either, but within an assembled spliceosomal complex hPrp28 gains ATP-binding activity. In order to understand the structural basis for the ATP-binding deficiency of isolated hPrp28, the crystal structure of hPrp28ΔN was determined at 2.0 Å resolution. In the crystal the helicase domain adopts a wide-open conformation, as the two RecA-like domains are extraordinarily displaced from the productive ATPase conformation. Binding of ATP is hindered by a closed conformation of themore » P-loop, which occupies the space required for the γ-phosphate of ATP.« less
Authors:
 [1] ;  [2] ; ;  [1] ;  [2] ;  [1]
  1. Georg-August-University Göttingen, Justus-von-Liebig Weg 11, 37077 Göttingen (Germany)
  2. Max-Planck-Institute for Biophysical Chemistry, Am Fassberg, 37077 Göttingen (Germany)
Publication Date:
OSTI Identifier:
22347765
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography; Journal Volume: 70; Journal Issue: Pt 6; Other Information: PMCID: PMC4051504; PMID: 24914973; PUBLISHER-ID: dw5092; OAI: oai:pubmedcentral.nih.gov:4051504; Copyright (c) Möhlmann et al. 2014; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
Denmark
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTAL STRUCTURE; CRYSTALS; FUNCTIONAL ANALYSIS; LENGTH; PHOSPHATES; PROTEINS; RESOLUTION