Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
Mol Cell Biol. 1997 Apr; 17(4): 2158–2165.
PMCID: PMC232064

Characterization of a beta-actin mRNA zipcode-binding protein.


Localization of beta-actin mRNA to the leading edge of fibroblasts requires the presence of conserved elements in the 3' untranslated region of the mRNA, including a 54-nucleotide element which has been termed the "zipcode" (E. Kislauskis, X. Zhu, and R. H. Singer, J. Cell Biol. 127:441-451, 1994). In order to identify proteins which bind to the zipcode and possibly play a role in localization, we performed band-shift mobility assays, UV cross-linking, and affinity purification experiments. A protein of 68 kDa was identified which binds to the proximal (to the coding region) half of the zipcode with high specificity (ZBP-1). Microsequencing provided unique peptide sequences of approximately 15 residues each. Degenerate primers corresponding to the codons derived from the peptides were synthesized and used for PCR amplification. Screening of a chicken cDNA library resulted in isolation of several clones providing a DNA sequence encoding a 67.7-kDa protein with regions homologous to several RNA-binding proteins, such as hnRNP E1 and E2, and with consensus mRNA recognition motif with RNP1 and 2 motifs and a putative REV-like nuclear export signal. Antipeptide antibodies were raised in rabbits which bound to ZBP-1 and coimmunoprecipitated proteins of 120 and 25 kDa. The 120-kDa protein was also obtained by affinity purification with the RNA zipcode sequence, along with a 53-kDa protein, but the 25-kDa protein appeared only in immunoprecipitations. Mutation of one of the conserved sequences within the zipcode, an ACACCC element in its proximal half, greatly reduced its protein binding and localization properties. These data suggest that the 68-kDa ZBP-1 we have isolated and cloned is an RNA-binding protein that functions within a complex to localize beta-actin mRNA.

Full Text

The Full Text of this article is available as a PDF (982K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ainger K, Avossa D, Morgan F, Hill SJ, Barry C, Barbarese E, Carson JH. Transport and localization of exogenous myelin basic protein mRNA microinjected into oligodendrocytes. J Cell Biol. 1993 Oct;123(2):431–441. [PMC free article] [PubMed]
  • Bassell GJ, Powers CM, Taneja KL, Singer RH. Single mRNAs visualized by ultrastructural in situ hybridization are principally localized at actin filament intersections in fibroblasts. J Cell Biol. 1994 Aug;126(4):863–876. [PMC free article] [PubMed]
  • Bassell GJ, Singer RH, Kosik KS. Association of poly(A) mRNA with microtubules in cultured neurons. Neuron. 1994 Mar;12(3):571–582. [PubMed]
  • Burd CG, Dreyfuss G. Conserved structures and diversity of functions of RNA-binding proteins. Science. 1994 Jul 29;265(5172):615–621. [PubMed]
  • Elisha Z, Havin L, Ringel I, Yisraeli JK. Vg1 RNA binding protein mediates the association of Vg1 RNA with microtubules in Xenopus oocytes. EMBO J. 1995 Oct 16;14(20):5109–5114. [PMC free article] [PubMed]
  • Ferrandon D, Elphick L, Nüsslein-Volhard C, St Johnston D. Staufen protein associates with the 3'UTR of bicoid mRNA to form particles that move in a microtubule-dependent manner. Cell. 1994 Dec 30;79(7):1221–1232. [PubMed]
  • Forristall C, Pondel M, Chen L, King ML. Patterns of localization and cytoskeletal association of two vegetally localized RNAs, Vg1 and Xcat-2. Development. 1995 Jan;121(1):201–208. [PubMed]
  • Gottlieb E. The 3' untranslated region of localized maternal messages contains a conserved motif involved in mRNA localization. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7164–7168. [PMC free article] [PubMed]
  • Hill MA, Schedlich L, Gunning P. Serum-induced signal transduction determines the peripheral location of beta-actin mRNA within the cell. J Cell Biol. 1994 Sep;126(5):1221–1229. [PMC free article] [PubMed]
  • Hill MA, Gunning P. Beta and gamma actin mRNAs are differentially located within myoblasts. J Cell Biol. 1993 Aug;122(4):825–832. [PMC free article] [PubMed]
  • Hoock TC, Newcomb PM, Herman IM. Beta actin and its mRNA are localized at the plasma membrane and the regions of moving cytoplasm during the cellular response to injury. J Cell Biol. 1991 Feb;112(4):653–664. [PMC free article] [PubMed]
  • Kim-Ha J, Kerr K, Macdonald PM. Translational regulation of oskar mRNA by bruno, an ovarian RNA-binding protein, is essential. Cell. 1995 May 5;81(3):403–412. [PubMed]
  • Kislauskis EH, Singer RH. Determinants of mRNA localization. Curr Opin Cell Biol. 1992 Dec;4(6):975–978. [PubMed]
  • Kislauskis EH, Li Z, Singer RH, Taneja KL. Isoform-specific 3'-untranslated sequences sort alpha-cardiac and beta-cytoplasmic actin messenger RNAs to different cytoplasmic compartments. J Cell Biol. 1993 Oct;123(1):165–172. [PMC free article] [PubMed]
  • Kislauskis EH, Zhu X, Singer RH. Sequences responsible for intracellular localization of beta-actin messenger RNA also affect cell phenotype. J Cell Biol. 1994 Oct;127(2):441–451. [PMC free article] [PubMed]
  • Kloc M, Etkin LD. Delocalization of Vg1 mRNA from the vegetal cortex in Xenopus oocytes after destruction of Xlsirt RNA. Science. 1994 Aug 19;265(5175):1101–1103. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Latham VM, Jr, Kislauskis EH, Singer RH, Ross AF. Beta-actin mRNA localization is regulated by signal transduction mechanisms. J Cell Biol. 1994 Sep;126(5):1211–1219. [PMC free article] [PubMed]
  • Lawrence JB, Singer RH. Intracellular localization of messenger RNAs for cytoskeletal proteins. Cell. 1986 May 9;45(3):407–415. [PubMed]
  • MacDonald PM, Leask A, Kerr K. exl protein specifically binds BLE1, a bicoid mRNA localization element, and is required for one phase of its activity. Proc Natl Acad Sci U S A. 1995 Nov 7;92(23):10787–10791. [PMC free article] [PubMed]
  • Mowry KL, Melton DA. Vegetal messenger RNA localization directed by a 340-nt RNA sequence element in Xenopus oocytes. Science. 1992 Feb 21;255(5047):991–994. [PubMed]
  • Murray MT, Krohne G, Franke WW. Different forms of soluble cytoplasmic mRNA binding proteins and particles in Xenopus laevis oocytes and embryos. J Cell Biol. 1991 Jan;112(1):1–11. [PMC free article] [PubMed]
  • Sundell CL, Singer RH. Actin mRNA localizes in the absence of protein synthesis. J Cell Biol. 1990 Dec;111(6 Pt 1):2397–2403. [PMC free article] [PubMed]
  • Sundell CL, Singer RH. Requirement of microfilaments in sorting of actin messenger RNA. Science. 1991 Sep 13;253(5025):1275–1277. [PubMed]
  • Wang S, Hazelrigg T. Implications for bcd mRNA localization from spatial distribution of exu protein in Drosophila oogenesis. Nature. 1994 Jun 2;369(6479):400–403. [PubMed]
  • Wang YL. Mobility of filamentous actin in living cytoplasm. J Cell Biol. 1987 Dec;105(6 Pt 1):2811–2816. [PMC free article] [PubMed]
  • Wilhelm JE, Vale RD. RNA on the move: the mRNA localization pathway. J Cell Biol. 1993 Oct;123(2):269–274. [PMC free article] [PubMed]
  • Yaffe D, Nudel U, Mayer Y, Neuman S. Highly conserved sequences in the 3' untranslated region of mRNAs coding for homologous proteins in distantly related species. Nucleic Acids Res. 1985 May 24;13(10):3723–3737. [PMC free article] [PubMed]
  • Yisraeli JK, Sokol S, Melton DA. A two-step model for the localization of maternal mRNA in Xenopus oocytes: involvement of microtubules and microfilaments in the translocation and anchoring of Vg1 mRNA. Development. 1990 Feb;108(2):289–298. [PubMed]

Articles from Molecular and Cellular Biology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...