Home LiteratureArticle Details
PMID: 12490714 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification of cellular mRNA targets for RNA-binding protein Sam68.

Nucleic acids research ·Vol. 30 ·No. 24 ·2002-12-15 ·Pages 5452-64

Itoh M, Haga I, Li QH, Fujisawa J

Abstract

Sam68 (Src-associated in mitosis, 68 kDa), a nuclear RNA-binding protein, has been postulated to play a role in cell-growth control as a modulator of signal transduction and activation of RNA metabolism. Although Sam68 was demonstrated to bind to the UAAA sequences in synthetic oligoribonucleotides and poly(U) homopolymers in vitro, the legitimate cellular mRNA target remained unclear. By using the differential display and cDNA-representational difference analysis techniques, followed by reverse transcription polymerase chain reaction of RNAs co-immunoprecipitated with Sam68 from a HeLa cell lysate, we identified 10 mRNA species that bind in vivo to Sam68 in an RNA-binding domain-dependent manner. Among them, the mRNA species for hnRNP A2/B1 and beta-actin were found to bind prominently in vivo as well as in vitro, suggesting the possible involvement of Sam68 in the post- transcriptional regulation of these genes. Mapping of the Sam68-binding sequence revealed that Sam68 associates with these mRNAs through different nucleotide motifs, UAAA for hnRNP A2/B1 mRNA and UUUUUU for beta-actin mRNA, and that both binding sequences must reside in a loop structure for recognition by Sam68. The results indicated that Sam68 recognizes both the UAAA motif and poly(U) sequences in vivo for binding to cellular target mRNAs.

MeSH Terms
3' Untranslated Regions/genetics,metabolism Actins/genetics Adaptor Proteins, Signal Transducing Base Sequence Binding Sites/genetics DNA, Complementary/genetics DNA-Binding Proteins Glutathione Transferase/genetics,metabolism HeLa Cells Heterogeneous-Nuclear Ribonucleoproteins/genetics Humans Molecular Sequence Data Mutation Nucleic Acid Conformation Protein Binding RNA, Messenger/chemistry,genetics,metabolism RNA-Binding Proteins/genetics,metabolism Recombinant Fusion Proteins/genetics,metabolism
Chemicals
3' Untranslated Regions Actins Adaptor Proteins, Signal Transducing DNA, Complementary DNA-Binding Proteins Heterogeneous-Nuclear Ribonucleoproteins KHDRBS1 protein, human RNA, Messenger RNA-Binding Proteins Recombinant Fusion Proteins Glutathione Transferase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Itoh Michiyasu
Department of Microbiology, Kansai Medical University, 10-15 Fumizono-cho, Moriguchi, Osaka 570-8506, Japan.
Haga Izumi
Li Qing-Hua
Fujisawa Jun-ichi
References (47)
47 references, click to expand
  1. The interaction and colocalization of Sam68 with the splicing-associated factor YT521-B in nuclear dots is regulated by the Src family kinase p59(fyn).
    Mol Biol Cell. 1999 Nov;10(11):3909-26 PMID: 10564280
  2. Identifying differences in mRNA expression by representational difference analysis of cDNA.
    Nucleic Acids Res. 1994 Dec 25;22(25):5640-8 PMID: 7838717
  3. All you wanted to know about SELEX.
    Mol Biol Rep. 1994;20(2):97-107 PMID: 7536299
  4. Structure and expression of the gene (HNRPA2B1) encoding the human hnRNP protein A2/B1.
    Genomics. 1995 Jan 20;25(2):365-71 PMID: 7789969
  5. Mutations in gld-1, a female germ cell-specific tumor suppressor gene in Caenorhabditis elegans, affect a conserved domain also found in Src-associated protein Sam68.
    Genes Dev. 1995 Jun 15;9(12):1491-504 PMID: 7601353
  6. Analysis of altered gene expression by differential display.
    Methods Enzymol. 1995;254:304-21 PMID: 8531695
  7. The quaking gene product necessary in embryogenesis and myelination combines features of RNA binding and signal transduction proteins.
    Nat Genet. 1996 Mar;12(3):260-5 PMID: 8589716
  8. Human protein Sam68 relocalization and interaction with poliovirus RNA polymerase in infected cells.
    Proc Natl Acad Sci U S A. 1996 Mar 19;93(6):2296-301 PMID: 8637866
  9. Mammalian splicing factor SF1 is encoded by variant cDNAs and binds to RNA.
    RNA. 1996 Aug;2(8):794-810 PMID: 8752089
  10. Identification of Itk/Tsk Src homology 3 domain ligands.
    J Biol Chem. 1996 Oct 11;271(41):25646-56 PMID: 8810341
  11. The role of a lymphoid-restricted, Grb2-like SH3-SH2-SH3 protein in T cell receptor signaling.
    J Biol Chem. 1997 Jan 10;272(2):894-902 PMID: 8995379
  12. A role for Sam68 in cell cycle progression antagonized by a spliced variant within the KH domain.
    J Biol Chem. 1997 Feb 7;272(6):3129-32 PMID: 9013542
  13. Interaction between Sam68 and Src family tyrosine kinases, Fyn and Lck, in T cell receptor signaling.
    J Biol Chem. 1997 Mar 7;272(10):6214-9 PMID: 9045636
  14. Characterization of a beta-actin mRNA zipcode-binding protein.
    Mol Cell Biol. 1997 Apr;17(4):2158-65 PMID: 9121465
  15. who encodes a KH RNA binding protein that functions in muscle development.
    Development. 1997 Apr;124(7):1323-32 PMID: 9118803
  16. The held out wings (how) Drosophila gene encodes a putative RNA-binding protein involved in the control of muscular and cardiac activity.
    Development. 1997 May;124(10):2087-98 PMID: 9169854
  17. Self-association of the single-KH-domain family members Sam68, GRP33, GLD-1, and Qk1: role of the KH domain.
    Mol Cell Biol. 1997 Oct;17(10):5707-18 PMID: 9315629
  18. Specificity and determinants of Sam68 RNA binding. Implications for the biological function of K homology domains.
    J Biol Chem. 1997 Oct 24;272(43):27274-80 PMID: 9341174
  19. STAR, a gene family involved in signal transduction and activation of RNA.
    Trends Genet. 1997 Dec;13(12):479-84 PMID: 9433137
  20. A family of Drosophila genes encoding quaking-related maxi-KH domains.
    Biochem Genet. 1998 Feb;36(1-2):51-64 PMID: 9562906
  21. Integrin binding and mechanical tension induce movement of mRNA and ribosomes to focal adhesions.
    Nature. 1998 Apr 16;392(6677):730-3 PMID: 9565036
  22. The identification of two Drosophila K homology domain proteins. Kep1 and SAM are members of the Sam68 family of GSG domain proteins.
    J Biol Chem. 1998 Nov 13;273(46):30122-30 PMID: 9804767
  23. Characterization of Sam68-like mammalian proteins SLM-1 and SLM-2: SLM-1 is a Src substrate during mitosis.
    Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):2710-5 PMID: 10077576
  24. T-STAR/ETOILE: a novel relative of SAM68 that interacts with an RNA-binding protein implicated in spermatogenesis.
    Hum Mol Genet. 1999 Jun;8(6):959-69 PMID: 10332027
  25. Inhibition of HIV replication by dominant negative mutants of Sam68, a functional homolog of HIV-1 Rev.
    Nat Med. 1999 Jun;5(6):635-42 PMID: 10371501
  26. Sam68, RNA helicase A and Tap cooperate in the post-transcriptional regulation of human immunodeficiency virus and type D retroviral mRNA.
    Oncogene. 2000 Jul 27;19(32):3570-5 PMID: 10951562
  27. Binding of an RNA trafficking response element to heterogeneous nuclear ribonucleoproteins A1 and A2.
    J Biol Chem. 2000 Dec 8;275(49):38286-95 PMID: 11024030
  28. Neoplastic transformation and tumorigenesis associated with sam68 protein deficiency in cultured murine fibroblasts.
    J Biol Chem. 2000 Dec 22;275(51):40195-201 PMID: 11032831
  29. Lipoxygenase mRNA silencing in erythroid differentiation: The 3'UTR regulatory complex controls 60S ribosomal subunit joining.
    Cell. 2001 Jan 26;104(2):281-90 PMID: 11207368
  30. Characterization of hnRNP A2 and B1 using monoclonal antibodies: intracellular distribution and metabolism through cell cycle.
    Immunol Lett. 2001 Feb 1;76(1):49-54 PMID: 11222913
  31. The STAR/GSG family protein rSLM-2 regulates the selection of alternative splice sites.
    J Biol Chem. 2001 Mar 23;276(12):8665-73 PMID: 11118435
  32. Heterogeneous nuclear ribonucleoprotein B1 as early cancer biomarker for occult cancer of human lungs and bronchial dysplasia.
    Cancer Res. 2001 Mar 1;61(5):1896-902 PMID: 11280744
  33. Identification of Sam68 arginine glycine-rich sequences capable of conferring nonspecific RNA binding to the GSG domain.
    J Biol Chem. 2001 Aug 17;276(33):30803-11 PMID: 11395494
  34. Stress-induced nuclear bodies are sites of accumulation of pre-mRNA processing factors.
    Mol Biol Cell. 2001 Nov;12(11):3502-14 PMID: 11694584
  35. Retardation of the G2-M phase progression on gene disruption of RNA binding protein Sam68 in the DT40 cell line.
    FEBS Lett. 2002 Aug 14;525(1-3):145-50 PMID: 12163178
  36. Cloning of a full-length complementary DNA for an Artemia salina glycine-rich protein. Structural relationship with RNA binding proteins.
    J Biol Chem. 1987 Oct 5;262(28):13377-80 PMID: 2443491
  37. Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase.
    Science. 1990 Aug 3;249(4968):505-10 PMID: 2200121
  38. Increased levels of p21ras-GTP and enhanced DNA synthesis accompany elevated tyrosyl phosphorylation of GAP-associated proteins, p190 and p62, in c-src overexpressors.
    Oncogene. 1993 Apr;8(4):959-67 PMID: 7681161
  39. hnRNP proteins and the biogenesis of mRNA.
    Annu Rev Biochem. 1993;62:289-321 PMID: 8352591
  40. An RNA-binding protein associated with Src through its SH2 and SH3 domains in mitosis.
    Nature. 1994 Apr 28;368(6474):867-71 PMID: 7512694
  41. A target for Src in mitosis.
    Nature. 1994 Apr 28;368(6474):871-4 PMID: 7512695
  42. Identification of Src, Fyn, and Lyn SH3-binding proteins: implications for a function of SH3 domains.
    Mol Cell Biol. 1994 Jul;14(7):4509-21 PMID: 7516469
  43. A protein that is highly related to GTPase-activating protein-associated p62 complexes with phospholipase C gamma.
    Mol Cell Biol. 1994 Aug;14(8):5466-73 PMID: 7518563
  44. Serum-induced signal transduction determines the peripheral location of beta-actin mRNA within the cell.
    J Cell Biol. 1994 Sep;126(5):1221-9 PMID: 8063859
  45. Sequences responsible for intracellular localization of beta-actin messenger RNA also affect cell phenotype.
    J Cell Biol. 1994 Oct;127(2):441-51 PMID: 7929587
  46. Association of p62, a multifunctional SH2- and SH3-domain-binding protein, with src family tyrosine kinases, Grb2, and phospholipase C gamma-1.
    Mol Cell Biol. 1995 Jan;15(1):186-97 PMID: 7799925
  47. Salpalpha and Salpbeta, growth-arresting homologs of Sam68.
    Gene. 1999 Nov 15;240(1):133-47 PMID: 10564820
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkf673
Published
2002-12-15
Pages
5452-64
Language
English
Region
England
NLM ID
0411011
PMCID
PMC140046
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/gkf673 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2