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PMID: 15198983 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

A new yeast PUF family protein, Puf6p, represses ASH1 mRNA translation and is required for its localization.

Genes & development ·Vol. 18 ·No. 12 ·2004-06-15 ·Pages 1452-65

Gu W, Deng Y, Zenklusen D, Singer RH

Abstract

In yeast Saccharomyces cerevisiae, Ash1p, a protein determinant for mating-type switching, is segregated within the daughter cell nucleus to establish asymmetry of HO expression. The accumulation of Ash1p results from ASH1 mRNA that is sorted as a ribonucleoprotein particle (mRNP or locasome) to the distal tip of the bud where translation occurs. To study the mechanism regulating ASH1 mRNA translation, we isolated the ASH1 locasome and characterized the associated proteins by MALDI-TOF. One of these proteins was Puf6p, a new member of the PUF family of highly conserved RNA-binding proteins such as Pumilio in Drosophila, responsible for translational repression, usually to effect asymmetric expression. Puf6p-bound PUF consensus sequences in the 3'UTR of ASH1 mRNA and repressed the translation of ASH1 mRNA both in vivo and in vitro. In the puf6 Delta strain, asymmetric localization of both Ash1p and ASH1 mRNA were significantly reduced. We propose that Puf6p is a protein that functions in the translational control of ASH1 mRNA, and this translational inhibition is necessary before localization can proceed.

MeSH Terms
3' Untranslated Regions DNA-Binding Proteins/biosynthesis,genetics Drosophila Proteins Gene Expression Regulation Protein Biosynthesis RNA, Messenger/genetics,metabolism RNA-Binding Proteins/physiology Repressor Proteins/biosynthesis,genetics,physiology Ribonucleoproteins/metabolism Saccharomyces cerevisiae Proteins/physiology Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Transcription Factors/biosynthesis,genetics
Chemicals
3' Untranslated Regions ASH1 protein, Drosophila ASH1 protein, S cerevisiae DNA-Binding Proteins Drosophila Proteins Puf6 protein, S cerevisiae RNA, Messenger RNA-Binding Proteins Repressor Proteins Ribonucleoproteins Saccharomyces cerevisiae Proteins Transcription Factors messenger ribonucleoprotein
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Gu Wei
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Deng Yingfeng
Zenklusen Daniel
Singer Robert H
References (44)
44 references, click to expand
  1. The myosin motor, Myo4p, binds Ash1 mRNA via the adapter protein, She3p.
    Proc Natl Acad Sci U S A. 2000 May 9;97(10):5273-8 PMID: 10792032
  2. Recruitment of Nanos to hunchback mRNA by Pumilio.
    Genes Dev. 1999 Oct 15;13(20):2704-12 PMID: 10541556
  3. She2p, a novel RNA-binding protein tethers ASH1 mRNA to the Myo4p myosin motor via She3p.
    EMBO J. 2000 Oct 16;19(20):5514-24 PMID: 11032818
  4. She2p is a novel RNA-binding protein that recruits the Myo4p-She3p complex to ASH1 mRNA.
    EMBO J. 2000 Dec 1;19(23):6592-601 PMID: 11101531
  5. Neuronal RNA localization and the cytoskeleton.
    Results Probl Cell Differ. 2001;34:41-56 PMID: 11288678
  6. The tandem affinity purification (TAP) method: a general procedure of protein complex purification.
    Methods. 2001 Jul;24(3):218-29 PMID: 11403571
  7. RNP localization and transport in yeast.
    Annu Rev Cell Dev Biol. 2001;17:297-310 PMID: 11687491
  8. A predominantly nuclear protein affecting cytoplasmic localization of beta-actin mRNA in fibroblasts and neurons.
    J Cell Biol. 2002 Jan 7;156(1):41-51 PMID: 11781334
  9. NANOS-3 and FBF proteins physically interact to control the sperm-oocyte switch in Caenorhabditis elegans.
    Curr Biol. 1999 Sep 23;9(18):1009-18 PMID: 10508609
  10. nos-1 and nos-2, two genes related to Drosophila nanos, regulate primordial germ cell development and survival in Caenorhabditis elegans.
    Development. 1999 Nov;126(21):4861-71 PMID: 10518502
  11. Post-transcriptional regulation through the HO 3'-UTR by Mpt5, a yeast homolog of Pumilio and FBF.
    EMBO J. 2001 Feb 1;20(3):552-61 PMID: 11157761
  12. A PUF family portrait: 3'UTR regulation as a way of life.
    Trends Genet. 2002 Mar;18(3):150-7 PMID: 11858839
  13. 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):1158-67 PMID: 11867544
  14. Modular recognition of RNA by a human pumilio-homology domain.
    Cell. 2002 Aug 23;110(4):501-12 PMID: 12202039
  15. Ribonucleoprotein-dependent localization of the yeast class V myosin Myo4p.
    J Cell Biol. 2002 Dec 23;159(6):971-82 PMID: 12499354
  16. Asymmetric sorting of ash1p in yeast results from inhibition of translation by localization elements in the mRNA.
    Mol Cell. 2002 Dec;10(6):1319-30 PMID: 12504008
  17. Widespread cytoplasmic mRNA transport in yeast: identification of 22 bud-localized transcripts using DNA microarray analysis.
    Proc Natl Acad Sci U S A. 2003 Sep 30;100(20):11429-34 PMID: 13679573
  18. RNA asymmetric distribution and daughter/mother differentiation in yeast.
    Curr Opin Microbiol. 2003 Dec;6(6):614-20 PMID: 14662358
  19. Extensive association of functionally and cytotopically related mRNAs with Puf family RNA-binding proteins in yeast.
    PLoS Biol. 2004 Mar;2(3):E79 PMID: 15024427
  20. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites.
    Gene. 1988 Dec 30;74(2):527-34 PMID: 3073106
  21. Isolation of yeast nuclei.
    Methods Enzymol. 1991;194:735-49 PMID: 2005821
  22. RNA regulatory elements mediate control of Drosophila body pattern by the posterior morphogen nanos.
    Cell. 1991 Nov 29;67(5):955-67 PMID: 1720354
  23. Binding of pumilio to maternal hunchback mRNA is required for posterior patterning in Drosophila embryos.
    Cell. 1995 Mar 10;80(5):747-56 PMID: 7889568
  24. Translational regulation of oskar mRNA by bruno, an ovarian RNA-binding protein, is essential.
    Cell. 1995 May 5;81(3):403-12 PMID: 7736592
  25. Mother cell-specific HO expression in budding yeast depends on the unconventional myosin myo4p and other cytoplasmic proteins.
    Cell. 1996 Mar 8;84(5):687-97 PMID: 8625407
  26. Identification of asymmetrically localized determinant, Ash1p, required for lineage-specific transcription of the yeast HO gene.
    Cell. 1996 Mar 8;84(5):711-22 PMID: 8625409
  27. Translational regulation of maternal mRNAs.
    Curr Opin Genet Dev. 1996 Aug;6(4):403-7 PMID: 8791537
  28. mRNA and cytoskeletal filaments.
    Curr Opin Cell Biol. 1997 Feb;9(1):109-15 PMID: 9013679
  29. Mating type switching in yeast controlled by asymmetric localization of ASH1 mRNA.
    Science. 1997 Jul 18;277(5324):383-7 PMID: 9219698
  30. Nanos and pumilio establish embryonic polarity in Drosophila by promoting posterior deadenylation of hunchback mRNA.
    Development. 1997 Aug;124(15):3015-23 PMID: 9247343
  31. Actin-dependent localization of an RNA encoding a cell-fate determinant in yeast.
    Nature. 1997 Sep 4;389(6646):90-3 PMID: 9288973
  32. A conserved RNA-binding protein that regulates sexual fates in the C. elegans hermaphrodite germ line.
    Nature. 1997 Dec 4;390(6659):477-84 PMID: 9393998
  33. Localization-dependent translation requires a functional interaction between the 5' and 3' ends of oskar mRNA.
    Genes Dev. 1998 Jun 1;12(11):1652-64 PMID: 9620852
  34. The Pumilio RNA-binding domain is also a translational regulator.
    Mol Cell. 1998 May;1(6):863-72 PMID: 9660969
  35. RNA localization in development.
    Annu Rev Biochem. 1998;67:335-94 PMID: 9759492
  36. Localization of ASH1 mRNA particles in living yeast.
    Mol Cell. 1998 Oct;2(4):437-45 PMID: 9809065
  37. The PUMILIO-RNA interaction: a single RNA-binding domain monomer recognizes a bipartite target sequence.
    Biochemistry. 1999 Jan 12;38(2):596-604 PMID: 9888799
  38. The travels of mRNAs through all cells large and small.
    FASEB J. 1999 Mar;13(3):447-54 PMID: 10064611
  39. Structural elements required for the localization of ASH1 mRNA and of a green fluorescent protein reporter particle in vivo.
    Curr Biol. 1999 Mar 25;9(6):333-6 PMID: 10209102
  40. ASH1 mRNA localization in yeast involves multiple secondary structural elements and Ash1 protein translation.
    Curr Biol. 1999 Mar 25;9(6):337-40 PMID: 10209099
  41. Association of the class V myosin Myo4p with a localised messenger RNA in budding yeast depends on She proteins.
    J Cell Sci. 1999 May;112 ( Pt 10):1511-8 PMID: 10212145
  42. Localization and anchoring of mRNA in budding yeast.
    Curr Biol. 1999 Jun 3;9(11):569-78 PMID: 10359695
  43. A generic protein purification method for protein complex characterization and proteome exploration.
    Nat Biotechnol. 1999 Oct;17(10):1030-2 PMID: 10504710
  44. Mechanisms of RNA localization and translational regulation.
    Curr Opin Genet Dev. 2000 Oct;10(5):476-88 PMID: 10980424
Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1189004
Published
2004-06-15
Pages
1452-65
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC423195
Subset
IM
Grants
NIGMS NIH HHS · R01 GM057071 · United States
NIGMS NIH HHS · GM 57071 · United States
External Links
PubMed source DOI: 10.1101/gad.1189004 PMC Full Text View journal details
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