• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biochemjBJ Latest papers and much more!
Biochem J. Jun 15, 2004; 380(Pt 3): 823–830.
PMCID: PMC1224212

Asc1p, a WD40-domain containing adaptor protein, is required for the interaction of the RNA-binding protein Scp160p with polysomes.

Abstract

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 Cys(1067) 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.

Full Text

The Full Text of this article is available as a PDF (505K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Lang BD, Fridovich-Keil JL. Scp160p, a multiple KH-domain protein, is a component of mRNP complexes in yeast. Nucleic Acids Res. 2000 Apr 1;28(7):1576–1584. [PMC free article] [PubMed]
  • Frey S, Pool M, Seedorf M. Scp160p, an RNA-binding, polysome-associated protein, localizes to the endoplasmic reticulum of Saccharomyces cerevisiae in a microtubule-dependent manner. J Biol Chem. 2001 May 11;276(19):15905–15912. [PubMed]
  • Lang BD, Li Am, Black-Brewster HD, Fridovich-Keil JL. The brefeldin A resistance protein Bfr1p is a component of polyribosome-associated mRNP complexes in yeast. Nucleic Acids Res. 2001 Jun 15;29(12):2567–2574. [PMC free article] [PubMed]
  • Kruse C, Grünweller A, Willkomm DK, Pfeiffer T, Hartmann RK, Müller PK. tRNA is entrapped in similar, but distinct, nuclear and cytoplasmic ribonucleoprotein complexes, both of which contain vigilin and elongation factor 1 alpha. Biochem J. 1998 Feb 1;329(Pt 3):615–621. [PMC free article] [PubMed]
  • Kruse C, Willkomm DK, Grünweller A, Vollbrandt T, Sommer S, Busch S, Pfeiffer T, Brinkmann J, Hartmann RK, Müller PK. Export and transport of tRNA are coupled to a multi-protein complex. Biochem J. 2000 Feb 15;346(Pt 1):107–115. [PMC free article] [PubMed]
  • Mendelsohn Bryce A, Li Ai-Min, Vargas Claudia A, Riehman Kristen, Watson Alice, Fridovich-Keil Judith L. Genetic and biochemical interactions between SCP160 and EAP1 in yeast. Nucleic Acids Res. 2003 Oct 15;31(20):5838–5847. [PMC free article] [PubMed]
  • Cosentino GP, Schmelzle T, Haghighat A, Helliwell SB, Hall MN, Sonenberg N. Eap1p, a novel eukaryotic translation initiation factor 4E-associated protein in Saccharomyces cerevisiae. Mol Cell Biol. 2000 Jul;20(13):4604–4613. [PMC free article] [PubMed]
  • Li Ai-Min, Watson Alice, Fridovich-Keil Judith L. Scp160p associates with specific mRNAs in yeast. Nucleic Acids Res. 2003 Apr 1;31(7):1830–1837. [PMC free article] [PubMed]
  • Guo Ming, Aston Christopher, Burchett Scott A, Dyke Christine, Fields Stanley, Rajarao S Johannes R, Uetz Peter, Wang Yuqi, Young Kathleen, Dohlman Henrik G. The yeast G protein alpha subunit Gpa1 transmits a signal through an RNA binding effector protein Scp160. Mol Cell. 2003 Aug;12(2):517–524. [PubMed]
  • Irie Kenji, Tadauchi Tomofumi, Takizawa Peter A, Vale Ronald D, Matsumoto Kunihiro, Herskowitz Ira. The Khd1 protein, which has three KH RNA-binding motifs, is required for proper localization of ASH1 mRNA in yeast. EMBO J. 2002 Mar 1;21(5):1158–1167. [PMC free article] [PubMed]
  • Wintersberger U, Kühne C, Karwan A. Scp160p, a new yeast protein associated with the nuclear membrane and the endoplasmic reticulum, is necessary for maintenance of exact ploidy. Yeast. 1995 Aug;11(10):929–944. [PubMed]
  • Cortés A, Huertas D, Fanti L, Pimpinelli S, Marsellach FX, Piña B, Azorín F. DDP1, a single-stranded nucleic acid-binding protein of Drosophila, associates with pericentric heterochromatin and is functionally homologous to the yeast Scp160p, which is involved in the control of cell ploidy. EMBO J. 1999 Jul 1;18(13):3820–3833. [PMC free article] [PubMed]
  • Sikorski RS, Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. [PMC free article] [PubMed]
  • Winston F, Dollard C, Ricupero-Hovasse SL. Construction of a set of convenient Saccharomyces cerevisiae strains that are isogenic to S288C. Yeast. 1995 Jan;11(1):53–55. [PubMed]
  • Chantrel Y, Gaisne M, Lions C, Verdière J. The transcriptional regulator Hap1p (Cyp1p) is essential for anaerobic or heme-deficient growth of Saccharomyces cerevisiae: Genetic and molecular characterization of an extragenic suppressor that encodes a WD repeat protein. Genetics. 1998 Feb;148(2):559–569. [PMC free article] [PubMed]
  • Knop M, Siegers K, Pereira G, Zachariae W, Winsor B, Nasmyth K, Schiebel E. Epitope tagging of yeast genes using a PCR-based strategy: more tags and improved practical routines. Yeast. 1999 Jul;15(10B):963–972. [PubMed]
  • Opitz Corinna, Di Cristina Manlio, Reiss Matthias, Ruppert Thomas, Crisanti Andrea, Soldati Dominique. Intramembrane cleavage of microneme proteins at the surface of the apicomplexan parasite Toxoplasma gondii. EMBO J. 2002 Apr 2;21(7):1577–1585. [PMC free article] [PubMed]
  • Link AJ, Eng J, Schieltz DM, Carmack E, Mize GJ, Morris DR, Garvik BM, Yates JR., 3rd Direct analysis of protein complexes using mass spectrometry. Nat Biotechnol. 1999 Jul;17(7):676–682. [PubMed]
  • Inada Toshifumi, Winstall Eric, Tarun Salvador Z, Jr, Yates John R, 3rd, Schieltz Dave, Sachs Alan B. One-step affinity purification of the yeast ribosome and its associated proteins and mRNAs. RNA. 2002 Jul;8(7):948–958. [PMC free article] [PubMed]
  • Shor Boris, Calaycay Jimmy, Rushbrook Julie, McLeod Maureen. Cpc2/RACK1 is a ribosome-associated protein that promotes efficient translation in Schizosaccharomyces pombe. J Biol Chem. 2003 Dec 5;278(49):49119–49128. [PubMed]
  • Brejning Jeanette, Jespersen Lene. Protein expression during lag phase and growth initiation in Saccharomyces cerevisiae. Int J Food Microbiol. 2002 May 5;75(1-2):27–38. [PubMed]
  • Stark H, Rodnina MV, Rinke-Appel J, Brimacombe R, Wintermeyer W, van Heel M. Visualization of elongation factor Tu on the Escherichia coli ribosome. Nature. 1997 Sep 25;389(6649):403–406. [PubMed]
  • Kruse C, Grünweller A, Notbohm H, Kügler S, Purschke WG, Müller PK. Evidence for a novel cytoplasmic tRNA-protein complex containing the KH-multidomain protein vigilin. Biochem J. 1996 Nov 15;320(Pt 1):247–252. [PMC free article] [PubMed]
  • Warner JR. The economics of ribosome biosynthesis in yeast. Trends Biochem Sci. 1999 Nov;24(11):437–440. [PubMed]
  • Hoffmann B, Mösch HU, Sattlegger E, Barthelmess IB, Hinnebusch A, Braus GH. The WD protein Cpc2p is required for repression of Gcn4 protein activity in yeast in the absence of amino-acid starvation. Mol Microbiol. 1999 Feb;31(3):807–822. [PubMed]
  • Neer EJ, Schmidt CJ, Nambudripad R, Smith TF. The ancient regulatory-protein family of WD-repeat proteins. Nature. 1994 Sep 22;371(6495):297–300. [PubMed]
  • Schechtman D, Mochly-Rosen D. Adaptor proteins in protein kinase C-mediated signal transduction. Oncogene. 2001 Oct 1;20(44):6339–6347. [PubMed]
  • McCahill Angela, Warwicker Jim, Bolger Graeme B, Houslay Miles D, Yarwood Stephen J. The RACK1 scaffold protein: a dynamic cog in cell response mechanisms. Mol Pharmacol. 2002 Dec;62(6):1261–1273. [PubMed]
  • Ron D, Chen CH, Caldwell J, Jamieson L, Orr E, Mochly-Rosen D. Cloning of an intracellular receptor for protein kinase C: a homolog of the beta subunit of G proteins. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):839–843. [PMC free article] [PubMed]
  • Ron D, Jiang Z, Yao L, Vagts A, Diamond I, Gordon A. Coordinated movement of RACK1 with activated betaIIPKC. J Biol Chem. 1999 Sep 17;274(38):27039–27046. [PubMed]
  • Angenstein Frank, Evans Anne M, Settlage Robert E, Moran Stewart T, Ling Shuo-Chien, Klintsova Anna Y, Shabanowitz Jeffrey, Hunt Donald F, Greenough William T. A receptor for activated C kinase is part of messenger ribonucleoprotein complexes associated with polyA-mRNAs in neurons. J Neurosci. 2002 Oct 15;22(20):8827–8837. [PubMed]
  • Ceci Marcello, Gaviraghi Cristina, Gorrini Chiara, Sala Leonardo A, Offenhäuser Nina, Marchisio Pier Carlo, Biffo Stefano. Release of eIF6 (p27BBP) from the 60S subunit allows 80S ribosome assembly. Nature. 2003 Dec 4;426(6966):579–584. [PubMed]
  • Liedtke Carole M, Yun C H Chris, Kyle Nicole, Wang Dandan. Protein kinase C epsilon-dependent regulation of cystic fibrosis transmembrane regulator involves binding to a receptor for activated C kinase (RACK1) and RACK1 binding to Na+/H+ exchange regulatory factor. J Biol Chem. 2002 Jun 21;277(25):22925–22933. [PubMed]
  • Hoffmann B, Wanke C, Lapaglia SK, Braus GH. c-Jun and RACK1 homologues regulate a control point for sexual development in Aspergillus nidulans. Mol Microbiol. 2000 Jul;37(1):28–41. [PubMed]
  • Goossens A, Dever TE, Pascual-Ahuir A, Serrano R. The protein kinase Gcn2p mediates sodium toxicity in yeast. J Biol Chem. 2001 Aug 17;276(33):30753–30760. [PubMed]
  • McLeod M, Shor B, Caporaso A, Wang W, Chen H, Hu L. Cpc2, a fission yeast homologue of mammalian RACK1 protein, interacts with Ran1 (Pat1) kinase To regulate cell cycle progression and meiotic development. Mol Cell Biol. 2000 Jun;20(11):4016–4027. [PMC free article] [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • MedGen
    MedGen
    Related information in MedGen
  • Nucleotide
    Nucleotide
    Published Nucleotide sequences
  • Protein
    Protein
    Published protein sequences
  • PubMed
    PubMed
    PubMed citations for these articles
  • Structure
    Structure
    Published 3D structures
  • Substance
    Substance
    PubChem Substance links