Scp160p interacts in an mRNA-dependent manner with translating ribosomes via multiple RNA-binding heterogeneous nuclear ribonucleoprotein K-homology (KH) domains. In the present study, we show by protein–protein cross-linking that Scp160p is in close proximity to translation elongation factor 1A and the WD40 (Trp-Asp 40)-repeat containing protein Asc1p at ribosomes. Analysis of a truncation mutant revealed that the C-terminus of Scp160p is essential for ribosome binding and that Cys1067 at the C-terminus of Scp160p is required to obtain these cross-links. The interaction of Scp160p with ribosomes depends on Asc1p. In fast-growing yeast cells, nearly all Asc1p is tightly bound to ribosomes, but it can also be present in a ribosome-free form depending on growth conditions. The functional homologue of Asc1p, mammalian RACK1 (receptor of activated C kinase), was previously characterized as an adaptor protein bridging activated signalling molecules with their substrates. Our results suggest that Scp160p connects specific mRNAs, ribosomes and a translation factor with an adaptor for signalling molecules. These interactions might regulate the translation activity of ribosomes programmed with specific mRNAs.
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Research Article|
June 15 2004
Asc1p, a WD40-domain containing adaptor protein, is required for the interaction of the RNA-binding protein Scp160p with polysomes
Sonja BAUM;
Sonja BAUM
Center of Molecular Biology at University of Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
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Margarethe BITTINS;
Margarethe BITTINS
Center of Molecular Biology at University of Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
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Steffen FREY;
Steffen FREY
Center of Molecular Biology at University of Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
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Matthias SEEDORF
Matthias SEEDORF
1
Center of Molecular Biology at University of Heidelberg (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
1To whom correspondence should be addressed (e-mail m.seedorf@zmbh.uni-heidelberg.de).
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Publisher: Portland Press Ltd
Received:
December 22 2003
Revision Received:
February 24 2004
Accepted:
March 11 2004
Accepted Manuscript online:
March 11 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2004
2004
Biochem J (2004) 380 (3): 823–830.
Article history
Received:
December 22 2003
Revision Received:
February 24 2004
Accepted:
March 11 2004
Accepted Manuscript online:
March 11 2004
Citation
Sonja BAUM, Margarethe BITTINS, Steffen FREY, Matthias SEEDORF; Asc1p, a WD40-domain containing adaptor protein, is required for the interaction of the RNA-binding protein Scp160p with polysomes. Biochem J 15 June 2004; 380 (3): 823–830. doi: https://doi.org/10.1042/bj20031962
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