Search tips
Search criteria 


Logo of jcellbiolHomeEditorsContactInstructions for Authors
J Cell Biol. 1993 October 1; 123(1): 165–172.
PMCID: PMC2119818

Isoform-specific 3'-untranslated sequences sort alpha-cardiac and beta- cytoplasmic actin messenger RNAs to different cytoplasmic compartments [published erratum appears in J Cell Biol 1993 Dec;123(6 Pt 2):following 1907]


We demonstrate that in differentiating myoblasts, the mRNAs encoding two actin isoforms, beta-cytoplasmic, and alpha-cardiac, can occupy different cytoplasmic compartments within the same cytoplasm. beta- actin mRNA is localized to the leading lamellae and alpha-actin mRNA is associated with a perinuclear compartment. This was revealed by co- hybridizing, in situ, fluorochrome-conjugated oligonucleotide probes specific for each isoform. To address the mechanism of isoform-specific mRNA localization, molecular chimeras were constructed by insertion of actin sequences between the Lac Z coding region and SV-40 3'UTR in a reporter plasmid. These constructs were transiently expressed in a mixed culture of embryonic fibroblasts, myoblasts and myotubes, beta- galactosidase activity within transfectants was revealed by a brief incubation with its substrate (X-gal). Since the blue-insoluble reaction product co-localized with the specific mRNAs expressed from each construct, it was used as a bioassay for mRNA localization. Transfectants were scored as either perinuclear, peripheral or nonlocalized with respect to the distribution of the blue product. The percentage of transfectants within those categories was quantitated as a function of the various constructs. This analysis revealed that for each actin mRNA its 3'UTR is necessary and sufficient to direct reporter transcripts to its appropriate compartment; beta-actin peripheral and alpha-actin perinuclear. In contrast, sequences from the 5'UTR through the coding region of either actin gene did not localize the blue product. Therefore, 3'UTR sequences play a key role in modulating the distribution of actin mRNAs in muscle cells. We propose that the mechanism of mRNA localization facilitates actin isoform sorting in the cytoplasm.

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.
  • An G, Hidaka K, Siminovitch L. Expression of bacterial beta-galactosidase in animal cells. Mol Cell Biol. 1982 Dec;2(12):1628–1632. [PMC free article] [PubMed]
  • Antin PB, Ordahl CP. Isolation and characterization of an avian myogenic cell line. Dev Biol. 1991 Jan;143(1):111–121. [PubMed]
  • Bergsma DJ, Chang KS, Schwartz RJ. Novel chicken actin gene: third cytoplasmic isoform. Mol Cell Biol. 1985 May;5(5):1151–1162. [PMC free article] [PubMed]
  • Chang KS, Zimmer WE, Jr, Bergsma DJ, Dodgson JB, Schwartz RJ. Isolation and characterization of six different chicken actin genes. Mol Cell Biol. 1984 Nov;4(11):2498–2508. [PMC free article] [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]
  • DeNofrio D, Hoock TC, Herman IM. Functional sorting of actin isoforms in microvascular pericytes. J Cell Biol. 1989 Jul;109(1):191–202. [PMC free article] [PubMed]
  • DePonti-Zilli L, Seiler-Tuyns A, Paterson BM. A 40-base-pair sequence in the 3' end of the beta-actin gene regulates beta-actin mRNA transcription during myogenesis. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1389–1393. [PubMed]
  • Gavis ER, Lehmann R. Localization of nanos RNA controls embryonic polarity. Cell. 1992 Oct 16;71(2):301–313. [PubMed]
  • Gorman CM, Merlino GT, Willingham MC, Pastan I, Howard BH. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777–6781. [PubMed]
  • Gunning P, Ponte P, Kedes L, Hickey RJ, Skoultchi AI. Expression of human cardiac actin in mouse L cells: a sarcomeric actin associates with a nonmuscle cytoskeleton. Cell. 1984 Mar;36(3):709–715. [PubMed]
  • Hayward LJ, Schwartz RJ. Sequential expression of chicken actin genes during myogenesis. J Cell Biol. 1986 Apr;102(4):1485–1493. [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]
  • Herman IM. Actin isoforms. Curr Opin Cell Biol. 1993 Feb;5(1):48–55. [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]
  • Jackson RJ. Cytoplasmic regulation of mRNA function: the importance of the 3' untranslated region. Cell. 1993 Jul 16;74(1):9–14. [PubMed]
  • Kislauskis EH, Singer RH. Determinants of mRNA localization. Curr Opin Cell Biol. 1992 Dec;4(6):975–978. [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]
  • Lim K, Chae CB. A simple assay for DNA transfection by incubation of the cells in culture dishes with substrates for beta-galactosidase. Biotechniques. 1989 Jun;7(6):576–579. [PubMed]
  • Lipshitz HD. Axis specification in the Drosophila embryo. Curr Opin Cell Biol. 1991 Dec;3(6):966–975. [PubMed]
  • Luby-Phelps K, Castle PE, Taylor DL, Lanni F. Hindered diffusion of inert tracer particles in the cytoplasm of mouse 3T3 cells. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4910–4913. [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]
  • McHugh KM, Crawford K, Lessard JL. A comprehensive analysis of the developmental and tissue-specific expression of the isoactin multigene family in the rat. Dev Biol. 1991 Dec;148(2):442–458. [PubMed]
  • McKenna N, Meigs JB, Wang YL. Identical distribution of fluorescently labeled brain and muscle actins in living cardiac fibroblasts and myocytes. J Cell Biol. 1985 Jan;100(1):292–296. [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]
  • Okabe S, Hirokawa N. Incorporation and turnover of biotin-labeled actin microinjected into fibroblastic cells: an immunoelectron microscopic study. J Cell Biol. 1989 Oct;109(4 Pt 1):1581–1595. [PMC free article] [PubMed]
  • Otey CA, Kalnoski MH, Bulinski JC. Immunolocalization of muscle and nonmuscle isoforms of actin in myogenic cells and adult skeletal muscle. Cell Motil Cytoskeleton. 1988;9(4):337–348. [PubMed]
  • Pardo JV, Pittenger MF, Craig SW. Subcellular sorting of isoactins: selective association of gamma actin with skeletal muscle mitochondria. Cell. 1983 Apr;32(4):1093–1103. [PubMed]
  • Paterson BM, Eldridge JD. alpha-Cardiac actin is the major sarcomeric isoform expressed in embryonic avian skeletal muscle. Science. 1984 Jun 29;224(4656):1436–1438. [PubMed]
  • Peltz SW, Jacobson A. mRNA stability: in trans-it. Curr Opin Cell Biol. 1992 Dec;4(6):979–983. [PubMed]
  • Peng I, Fischman DA. Post-translational incorporation of actin into myofibrils in vitro: evidence for isoform specificity. Cell Motil Cytoskeleton. 1991;20(2):158–168. [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]
  • Ralston E, Hall ZW. Restricted distribution of mRNA produced from a single nucleus in hybrid myotubes. J Cell Biol. 1992 Dec;119(5):1063–1068. [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]
  • Rubenstein PA. The functional importance of multiple actin isoforms. Bioessays. 1990 Jul;12(7):309–315. [PubMed]
  • Ruzicka DL, Schwartz RJ. Sequential activation of alpha-actin genes during avian cardiogenesis: vascular smooth muscle alpha-actin gene transcripts mark the onset of cardiomyocyte differentiation. J Cell Biol. 1988 Dec;107(6 Pt 2):2575–2586. [PMC free article] [PubMed]
  • Sadano H, Taniguchi S, Baba T. Newly identified type of beta actin reduces invasiveness of mouse B16-melanoma. FEBS Lett. 1990 Oct 1;271(1-2):23–27. [PubMed]
  • Sawtell NM, Lessard JL. Cellular distribution of smooth muscle actins during mammalian embryogenesis: expression of the alpha-vascular but not the gamma-enteric isoform in differentiating striated myocytes. J Cell Biol. 1989 Dec;109(6 Pt 1):2929–2937. [PMC free article] [PubMed]
  • Sawtell NM, Hartman AL, Lessard JL. Unique isoactins in the brush border of rat intestinal epithelial cells. Cell Motil Cytoskeleton. 1988;11(4):318–325. [PubMed]
  • Schevzov G, Lloyd C, Gunning P. High level expression of transfected beta- and gamma-actin genes differentially impacts on myoblast cytoarchitecture. J Cell Biol. 1992 May;117(4):775–785. [PMC free article] [PubMed]
  • Singer RH. The cytoskeleton and mRNA localization. Curr Opin Cell Biol. 1992 Feb;4(1):15–19. [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]
  • St Johnston D, Nüsslein-Volhard C. The origin of pattern and polarity in the Drosophila embryo. Cell. 1992 Jan 24;68(2):201–219. [PubMed]
  • Taneja K, Singer RH. Use of oligodeoxynucleotide probes for quantitative in situ hybridization to actin mRNA. Anal Biochem. 1987 Nov 1;166(2):389–398. [PubMed]
  • Taneja KL, Singer RH. Detection and localization of actin mRNA isoforms in chicken muscle cells by in situ hybridization using biotinated oligonucleotide probes. J Cell Biochem. 1990 Dec;44(4):241–252. [PubMed]
  • Taneja KL, Lifshitz LM, Fay FS, Singer RH. Poly(A) RNA codistribution with microfilaments: evaluation by in situ hybridization and quantitative digital imaging microscopy. J Cell Biol. 1992 Dec;119(5):1245–1260. [PMC free article] [PubMed]
  • Theriot JA, Mitchison TJ. Actin microfilament dynamics in locomoting cells. Nature. 1991 Jul 11;352(6331):126–131. [PubMed]
  • Vandekerckhove J, Weber K. At least six different actins are expressed in a higher mammal: an analysis based on the amino acid sequence of the amino-terminal tryptic peptide. J Mol Biol. 1978 Dec 25;126(4):783–802. [PubMed]
  • Vandekerckhove J, Weber K. Chordate muscle actins differ distinctly from invertebrate muscle actins. The evolution of the different vertebrate muscle actins. J Mol Biol. 1984 Nov 5;179(3):391–413. [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]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press