Abstract
3T3-L1 preadipocytes ectopically expressing the mammalian RNA-binding protein Hel-N1 expressed up to 10-fold more glucose transporter (GLUT1) protein and exhibited elevated rates of basal glucose uptake. Hel-N1 is a member of the ELAV-like family of proteins associated with the induction and maintenance of differentiation in various species. ELAV proteins are known to bind in vitro to short stretches of uridylates in the 3' untranslated regions (3'UTRs) of unstable mRNAs encoding growth-regulatory proteins involved in transcription and signal transduction. GLUT1 mRNA also contains a large 3'UTR with a U-rich region that binds specifically to Hel-N1 in vitro. Analysis of the altered GLUT1 expression at the translational and posttranscriptional levels suggested a mechanism involving both mRNA stabilization and accelerated formation of translation initiation complexes. These findings are consistent with the hypothesis that the Hel-N1 family of proteins modulate gene expression at the level of mRNA in the cytoplasm.
Full Text
The Full Text of this article is available as a PDF (688.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Casey J. L., Hentze M. W., Koeller D. M., Caughman S. W., Rouault T. A., Klausner R. D., Harford J. B. Iron-responsive elements: regulatory RNA sequences that control mRNA levels and translation. Science. 1988 May 13;240(4854):924–928. doi: 10.1126/science.2452485. [DOI] [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- Chung S., Jiang L., Cheng S., Furneaux H. Purification and properties of HuD, a neuronal RNA-binding protein. J Biol Chem. 1996 May 10;271(19):11518–11524. doi: 10.1074/jbc.271.19.11518. [DOI] [PubMed] [Google Scholar]
- Cleveland D. W., Lopata M. A., MacDonald R. J., Cowan N. J., Rutter W. J., Kirschner M. W. 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. doi: 10.1016/0092-8674(80)90238-x. [DOI] [PubMed] [Google Scholar]
- Cornelius P., MacDougald O. A., Lane M. D. Regulation of adipocyte development. Annu Rev Nutr. 1994;14:99–129. doi: 10.1146/annurev.nu.14.070194.000531. [DOI] [PubMed] [Google Scholar]
- Cornelius P., Marlowe M., Call K., Pekala P. H. Regulation of glucose transport as well as glucose transporter and immediate early gene expression in 3T3-L1 preadipocytes by 8-bromo-cAMP. J Cell Physiol. 1991 Feb;146(2):298–308. doi: 10.1002/jcp.1041460215. [DOI] [PubMed] [Google Scholar]
- Cornelius P., Marlowe M., Lee M. D., Pekala P. H. The growth factor-like effects of tumor necrosis factor-alpha. Stimulation of glucose transport activity and induction of glucose transporter and immediate early gene expression in 3T3-L1 preadipocytes. J Biol Chem. 1990 Nov 25;265(33):20506–20516. [PubMed] [Google Scholar]
- Cullen B. R. Trans-activation of human immunodeficiency virus occurs via a bimodal mechanism. Cell. 1986 Sep 26;46(7):973–982. doi: 10.1016/0092-8674(86)90696-3. [DOI] [PubMed] [Google Scholar]
- Frost S. C., Lane M. D. Evidence for the involvement of vicinal sulfhydryl groups in insulin-activated hexose transport by 3T3-L1 adipocytes. J Biol Chem. 1985 Mar 10;260(5):2646–2652. [PubMed] [Google Scholar]
- Fushiki T., Wells J. A., Tapscott E. B., Dohm G. L. Changes in glucose transporters in muscle in response to exercise. Am J Physiol. 1989 May;256(5 Pt 1):E580–E587. doi: 10.1152/ajpendo.1989.256.5.E580. [DOI] [PubMed] [Google Scholar]
- Gallie D. R., Walbot V. RNA pseudoknot domain of tobacco mosaic virus can functionally substitute for a poly(A) tail in plant and animal cells. Genes Dev. 1990 Jul;4(7):1149–1157. doi: 10.1101/gad.4.7.1149. [DOI] [PubMed] [Google Scholar]
- Gao F. B., Carson C. C., Levine T., Keene J. D. Selection of a subset of mRNAs from combinatorial 3' untranslated region libraries using neuronal RNA-binding protein Hel-N1. Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):11207–11211. doi: 10.1073/pnas.91.23.11207. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gao F. B., Keene J. D. Hel-N1/Hel-N2 proteins are bound to poly(A)+ mRNA in granular RNP structures and are implicated in neuronal differentiation. J Cell Sci. 1996 Mar;109(Pt 3):579–589. doi: 10.1242/jcs.109.3.579. [DOI] [PubMed] [Google Scholar]
- Good P. J. A conserved family of elav-like genes in vertebrates. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4557–4561. doi: 10.1073/pnas.92.10.4557. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goodwin E. B., Okkema P. G., Evans T. C., Kimble J. Translational regulation of tra-2 by its 3' untranslated region controls sexual identity in C. elegans. Cell. 1993 Oct 22;75(2):329–339. doi: 10.1016/0092-8674(93)80074-o. [DOI] [PubMed] [Google Scholar]
- Kaestner K. H., Christy R. J., McLenithan J. C., Braiterman L. T., Cornelius P., Pekala P. H., Lane M. D. Sequence, tissue distribution, and differential expression of mRNA for a putative insulin-responsive glucose transporter in mouse 3T3-L1 adipocytes. Proc Natl Acad Sci U S A. 1989 May;86(9):3150–3154. doi: 10.1073/pnas.86.9.3150. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaestner K. H., Flores-Riveros J. R., McLenithan J. C., Janicot M., Lane M. D. Transcriptional repression of the mouse insulin-responsive glucose transporter (GLUT4) gene by cAMP. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1933–1937. doi: 10.1073/pnas.88.5.1933. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kern M., Wells J. A., Stephens J. M., Elton C. W., Friedman J. E., Tapscott E. B., Pekala P. H., Dohm G. L. Insulin responsiveness in skeletal muscle is determined by glucose transporter (Glut4) protein level. Biochem J. 1990 Sep 1;270(2):397–400. doi: 10.1042/bj2700397. [DOI] [PMC free article] [PubMed] [Google Scholar]
- King P. H., Levine T. D., Fremeau R. T., Jr, Keene J. D. Mammalian homologs of Drosophila ELAV localized to a neuronal subset can bind in vitro to the 3' UTR of mRNA encoding the Id transcriptional repressor. J Neurosci. 1994 Apr;14(4):1943–1952. doi: 10.1523/JNEUROSCI.14-04-01943.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klip A., Walker D. The glucose transport system of muscle plasma membranes: characterization by means of [3H]cytochalasin B binding. Arch Biochem Biophys. 1983 Feb 15;221(1):175–187. doi: 10.1016/0003-9861(83)90134-0. [DOI] [PubMed] [Google Scholar]
- Kruys V. I., Wathelet M. G., Huez G. A. Identification of a translation inhibitory element (TIE) in the 3' untranslated region of the human interferon-beta mRNA. Gene. 1988 Dec 10;72(1-2):191–200. doi: 10.1016/0378-1119(88)90144-8. [DOI] [PubMed] [Google Scholar]
- Kumagai A. K., Dwyer K. J., Pardridge W. M. Differential glycosylation of the GLUT1 glucose transporter in brain capillaries and choroid plexus. Biochim Biophys Acta. 1994 Jul 13;1193(1):24–30. doi: 10.1016/0005-2736(94)90328-x. [DOI] [PubMed] [Google Scholar]
- Levine T. D., Gao F., King P. H., Andrews L. G., Keene J. D. Hel-N1: an autoimmune RNA-binding protein with specificity for 3' uridylate-rich untranslated regions of growth factor mRNAs. Mol Cell Biol. 1993 Jun;13(6):3494–3504. doi: 10.1128/mcb.13.6.3494. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Long S. D., Pekala P. H. Regulation of GLUT4 mRNA stability by tumor necrosis factor-alpha: alterations in both protein binding to the 3' untranslated region and initiation of translation. Biochem Biophys Res Commun. 1996 Mar 27;220(3):949–953. doi: 10.1006/bbrc.1996.0512. [DOI] [PubMed] [Google Scholar]
- MacDougald O. A., Lane M. D. Transcriptional regulation of gene expression during adipocyte differentiation. Annu Rev Biochem. 1995;64:345–373. doi: 10.1146/annurev.bi.64.070195.002021. [DOI] [PubMed] [Google Scholar]
- Maher F., Vannucci S. J., Simpson I. A. Glucose transporter proteins in brain. FASEB J. 1994 Oct;8(13):1003–1011. doi: 10.1096/fasebj.8.13.7926364. [DOI] [PubMed] [Google Scholar]
- Masumi A., Akamatsu Y., Kitagawa T. Modulation of the synthesis and glycosylation of the glucose transporter protein by transforming growth factor-beta 1 in Swiss 3T3 fibroblasts. Biochim Biophys Acta. 1993 Feb 9;1145(2):227–234. doi: 10.1016/0005-2736(93)90293-9. [DOI] [PubMed] [Google Scholar]
- McCarthy J. E., Kollmus H. Cytoplasmic mRNA-protein interactions in eukaryotic gene expression. Trends Biochem Sci. 1995 May;20(5):191–197. doi: 10.1016/s0968-0004(00)89006-4. [DOI] [PubMed] [Google Scholar]
- McGowan K. M., Long S. D., Pekala P. H. Glucose transporter gene expression: regulation of transcription and mRNA stability. Pharmacol Ther. 1995 Jun;66(3):465–505. doi: 10.1016/0163-7258(95)00007-4. [DOI] [PubMed] [Google Scholar]
- McMahon R. J., Frost S. C. Nutrient control of GLUT1 processing and turnover in 3T3-L1 adipocytes. J Biol Chem. 1995 May 19;270(20):12094–12099. doi: 10.1074/jbc.270.20.12094. [DOI] [PubMed] [Google Scholar]
- Murata Y., Wharton R. P. Binding of pumilio to maternal hunchback mRNA is required for posterior patterning in Drosophila embryos. Cell. 1995 Mar 10;80(5):747–756. doi: 10.1016/0092-8674(95)90353-4. [DOI] [PubMed] [Google Scholar]
- Ostareck-Lederer A., Ostareck D. H., Standart N., Thiele B. J. Translation of 15-lipoxygenase mRNA is inhibited by a protein that binds to a repeated sequence in the 3' untranslated region. EMBO J. 1994 Mar 15;13(6):1476–1481. doi: 10.1002/j.1460-2075.1994.tb06402.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pandey N. B., Marzluff W. F. The stem-loop structure at the 3' end of histone mRNA is necessary and sufficient for regulation of histone mRNA stability. Mol Cell Biol. 1987 Dec;7(12):4557–4559. doi: 10.1128/mcb.7.12.4557. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pekala P., Kawakami M., Vine W., Lane M. D., Cerami A. Studies of insulin resistance in adipocytes induced by macrophage mediator. J Exp Med. 1983 Apr 1;157(4):1360–1365. doi: 10.1084/jem.157.4.1360. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Query C. C., Bentley R. C., Keene J. D. A common RNA recognition motif identified within a defined U1 RNA binding domain of the 70K U1 snRNP protein. Cell. 1989 Apr 7;57(1):89–101. doi: 10.1016/0092-8674(89)90175-x. [DOI] [PubMed] [Google Scholar]
- Romac J. M., Graff D. H., Keene J. D. The U1 small nuclear ribonucleoprotein (snRNP) 70K protein is transported independently of U1 snRNP particles via a nuclear localization signal in the RNA-binding domain. Mol Cell Biol. 1994 Jul;14(7):4662–4670. doi: 10.1128/mcb.14.7.4662. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shaw G., Kamen R. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell. 1986 Aug 29;46(5):659–667. doi: 10.1016/0092-8674(86)90341-7. [DOI] [PubMed] [Google Scholar]
- Stephens J. M., Pekala P. H. Transcriptional repression of the C/EBP-alpha and GLUT4 genes in 3T3-L1 adipocytes by tumor necrosis factor-alpha. Regulations is coordinate and independent of protein synthesis. J Biol Chem. 1992 Jul 5;267(19):13580–13584. [PubMed] [Google Scholar]
- Stephens J. M., Pekala P. H. Transcriptional repression of the GLUT4 and C/EBP genes in 3T3-L1 adipocytes by tumor necrosis factor-alpha. J Biol Chem. 1991 Nov 15;266(32):21839–21845. [PubMed] [Google Scholar]
- Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
- Tebbey P. W., McGowan K. M., Stephens J. M., Buttke T. M., Pekala P. H. Arachidonic acid down-regulates the insulin-dependent glucose transporter gene (GLUT4) in 3T3-L1 adipocytes by inhibiting transcription and enhancing mRNA turnover. J Biol Chem. 1994 Jan 7;269(1):639–644. [PubMed] [Google Scholar]
- Tsai D. E., Kenan D. J., Keene J. D. In vitro selection of an RNA epitope immunologically cross-reactive with a peptide. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):8864–8868. doi: 10.1073/pnas.89.19.8864. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yang J., Clark A. E., Kozka I. J., Cushman S. W., Holman G. D. Development of an intracellular pool of glucose transporters in 3T3-L1 cells. J Biol Chem. 1992 May 25;267(15):10393–10399. [PubMed] [Google Scholar]