PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
 
J Cell Biol. 1994 October 2; 127(2): 441–451.
PMCID: PMC2120214

Sequences responsible for intracellular localization of beta-actin messenger RNA also affect cell phenotype

Abstract

We have characterized the structure and function of RNA sequences that direct beta-cytoplasmic actin mRNA to the cell periphery were mapped to two segments of 3'-untranslated region by expression of LacZ/beta-actin chimeric mRNAs in chicken embryo fibroblasts (CEFs). A 54-nt segment, the "RNA zipcode," and a homologous but less active 43-nt segment each localized beta-galactosidase activity to the leading lamellae. This zipcode contains the full activity, and mutations or deletions within it reduce, but do not eliminate, its activity, indicating that several motifs contribute to the activity. Two of these motifs, when multimerized, can regenerate almost full activity. These sequences are highly conserved in evolution, since the human beta-actin zipcode, positioned identically in the 3'UTR localizes equally well in chicken cells. Complementary phosphorothioate oligonucleotides against the zipcode delocalized endogenous beta-actin mRNA, whereas those complementary to the region just outside the zipcode, or sense oligonucleotides, did not. Actin mRNA or protein levels were unaffected by the antisense treatments, but a dramatic change in lamellipodia structure, and actin stress fiber organization was observed using the same antizipcode oligonucleotides which delocalized the mRNA. Hence, discrete 3'UTR sequences direct beta-actin isoform synthesis to the leading lamellae and affect cell morphology, presumably through the actin cytoskeleton.

Full Text

The Full Text of this article is available as a PDF (5.1M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Carlier MF. Actin: protein structure and filament dynamics. J Biol Chem. 1991 Jan 5;266(1):1–4. [PubMed]
  • Cheung HK, Serano TL, Cohen RS. Evidence for a highly selective RNA transport system and its role in establishing the dorsoventral axis of the Drosophila egg. Development. 1992 Mar;114(3):653–661. [PubMed]
  • Cleveland DW, Lopata MA, MacDonald RJ, Cowan NJ, Rutter WJ, Kirschner MW. Number and evolutionary conservation of alpha- and beta-tubulin and cytoplasmic beta- and gamma-actin genes using specific cloned cDNA probes. Cell. 1980 May;20(1):95–105. [PubMed]
  • Cooper JA. The role of actin polymerization in cell motility. Annu Rev Physiol. 1991;53:585–605. [PubMed]
  • Dalby B, Glover DM. Discrete sequence elements control posterior pole accumulation and translational repression of maternal cyclin B RNA in Drosophila. EMBO J. 1993 Mar;12(3):1219–1227. [PubMed]
  • Davis I, Ish-Horowicz D. Apical localization of pair-rule transcripts requires 3' sequences and limits protein diffusion in the Drosophila blastoderm embryo. Cell. 1991 Nov 29;67(5):927–940. [PubMed]
  • Drubin DG. Actin and actin-binding proteins in yeast. Cell Motil Cytoskeleton. 1990;15(1):7–11. [PubMed]
  • Drubin DG, Mulholland J, Zhu ZM, Botstein D. Homology of a yeast actin-binding protein to signal transduction proteins and myosin-I. Nature. 1990 Jan 18;343(6255):288–290. [PubMed]
  • Evans TC, Crittenden SL, Kodoyianni V, Kimble J. Translational control of maternal glp-1 mRNA establishes an asymmetry in the C. elegans embryo. Cell. 1994 Apr 22;77(2):183–194. [PubMed]
  • Farwell AP, Lynch RM, Okulicz WC, Comi AM, Leonard JL. The actin cytoskeleton mediates the hormonally regulated translocation of type II iodothyronine 5'-deiodinase in astrocytes. J Biol Chem. 1990 Oct 25;265(30):18546–18553. [PubMed]
  • Gavis ER, Lehmann R. Localization of nanos RNA controls embryonic polarity. Cell. 1992 Oct 16;71(2):301–313. [PubMed]
  • Haile DJ, Rouault TA, Tang CK, Chin J, Harford JB, Klausner RD. Reciprocal control of RNA-binding and aconitase activity in the regulation of the iron-responsive element binding protein: role of the iron-sulfur cluster. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7536–7540. [PubMed]
  • Hesketh JE, Pryme IF. Interaction between mRNA, ribosomes and the cytoskeleton. Biochem J. 1991 Jul 1;277(Pt 1):1–10. [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]
  • Hunt AN, Normand CS, Postle AD. CTP:cholinephosphate cytidylyltransferase in human and rat lung: association in vitro with cytoskeletal actin. Biochim Biophys Acta. 1990 Mar 12;1043(1):19–26. [PubMed]
  • Jackson RJ. Cytoplasmic regulation of mRNA function: the importance of the 3' untranslated region. Cell. 1993 Jul 16;74(1):9–14. [PubMed]
  • Kim-Ha J, Webster PJ, Smith JL, Macdonald PM. Multiple RNA regulatory elements mediate distinct steps in localization of oskar mRNA. Development. 1993 Sep;119(1):169–178. [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]
  • Klausner RD, Rouault TA, Harford JB. Regulating the fate of mRNA: the control of cellular iron metabolism. Cell. 1993 Jan 15;72(1):19–28. [PubMed]
  • Knull HR, Walsh JL. Association of glycolytic enzymes with the cytoskeleton. Curr Top Cell Regul. 1992;33:15–30. [PubMed]
  • Kuznetsov SA, Langford GM, Weiss DG. Actin-dependent organelle movement in squid axoplasm. Nature. 1992 Apr 23;356(6371):722–725. [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]
  • Lawrence JB, Taneja K, Singer RH. Temporal resolution and sequential expression of muscle-specific genes revealed by in situ hybridization. Dev Biol. 1989 May;133(1):235–246. [PubMed]
  • Leavitt J, Ng SY, Aebi U, Varma M, Latter G, Burbeck S, Kedes L, Gunning P. Expression of transfected mutant beta-actin genes: alterations of cell morphology and evidence for autoregulation in actin pools. Mol Cell Biol. 1987 Jul;7(7):2457–2466. [PMC free article] [PubMed]
  • Lloyd C, Gunning P. Noncoding regions of the gamma-actin gene influence the impact of the gene on myoblast morphology. J Cell Biol. 1993 Apr;121(1):73–82. [PMC free article] [PubMed]
  • MacDonald PM. bicoid mRNA localization signal: phylogenetic conservation of function and RNA secondary structure. Development. 1990 Sep;110(1):161–171. [PubMed]
  • Macdonald PM, Struhl G. cis-acting sequences responsible for anterior localization of bicoid mRNA in Drosophila embryos. Nature. 1988 Dec 8;336(6199):595–598. [PubMed]
  • Macdonald PM, Kerr K, Smith JL, Leask A. RNA regulatory element BLE1 directs the early steps of bicoid mRNA localization. Development. 1993 Aug;118(4):1233–1243. [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]
  • Negrutskii BS, Deutscher MP. Channeling of aminoacyl-tRNA for protein synthesis in vivo. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4991–4995. [PubMed]
  • Otey CA, Kalnoski MH, Bulinski JC. Identification and quantification of actin isoforms in vertebrate cells and tissues. J Cell Biochem. 1987 Jun;34(2):113–124. [PubMed]
  • Peltz SW, Jacobson A. mRNA stability: in trans-it. Curr Opin Cell Biol. 1992 Dec;4(6):979–983. [PubMed]
  • Pokrywka NJ, Stephenson EC. Microtubules mediate the localization of bicoid RNA during Drosophila oogenesis. Development. 1991 Sep;113(1):55–66. [PubMed]
  • Ponte P, Gunning P, Blau H, Kedes L. Human actin genes are single copy for alpha-skeletal and alpha-cardiac actin but multicopy for beta- and gamma-cytoskeletal genes: 3' untranslated regions are isotype specific but are conserved in evolution. Mol Cell Biol. 1983 Oct;3(10):1783–1791. [PMC free article] [PubMed]
  • Rastinejad F, Blau HM. Genetic complementation reveals a novel regulatory role for 3' untranslated regions in growth and differentiation. Cell. 1993 Mar 26;72(6):903–917. [PubMed]
  • Rastinejad F, Conboy MJ, Rando TA, Blau HM. Tumor suppression by RNA from the 3' untranslated region of alpha-tropomyosin. Cell. 1993 Dec 17;75(6):1107–1117. [PubMed]
  • Särndahl E, Lindroth M, Bengtsson T, Fällman M, Gustavsson J, Stendahl O, Andersson T. Association of ligand-receptor complexes with actin filaments in human neutrophils: a possible regulatory role for a G-protein. J Cell Biol. 1989 Dec;109(6 Pt 1):2791–2799. [PMC free article] [PubMed]
  • Singer RH. The cytoskeleton and mRNA localization. Curr Opin Cell Biol. 1992 Feb;4(1):15–19. [PubMed]
  • Singer RH. RNA zipcodes for cytoplasmic addresses. Curr Biol. 1993 Oct 1;3(10):719–721. [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]
  • Theurkauf WE, Smiley S, Wong ML, Alberts BM. Reorganization of the cytoskeleton during Drosophila oogenesis: implications for axis specification and intercellular transport. Development. 1992 Aug;115(4):923–936. [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]
  • Wharton RP, Struhl G. RNA regulatory elements mediate control of Drosophila body pattern by the posterior morphogen nanos. Cell. 1991 Nov 29;67(5):955–967. [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]
  • Yang F, Demma M, Warren V, Dharmawardhane S, Condeelis J. Identification of an actin-binding protein from Dictyostelium as elongation factor 1a. Nature. 1990 Oct 4;347(6292):494–496. [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 The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press