Abstract
The DNA sequence between position +36 and -1907 of the murine myelin basic protein gene contains the enhancer and promoter elements necessary for abundant and cell specific expression in transgenic mice. Surprisingly, the pattern of expression promoted by this DNA fragment is a subset of that exhibited by the endogenous myelin basic protein (MBP) gene. Fusion genes prepared with this promoter/enhancer and a Lac Z reporter gene are expressed only in oligodendrocytes and not in Schwann cells, whereas the endogenous MBP gene is expressed in both cell types. The level of transgene expression measured by nuclear run- on experiments is very substantial and rivals that of the endogenous MBP gene. Furthermore, this 1.9-kb DNA fragment directs transcription on the same (or very similar) developmental schedule as the endogenous gene. These results indicate that the MBP promoter/enhancer sequences are at least tripartite: a core promoter, the oligodendrocyte enhancer elements, and a third component that either expands the specificity of the oligodendrocyte enhancer to include Schwann cells or acts independently to specifically stimulate transcription in Schwann cells.
Full Text
The Full Text of this article is available as a PDF (6.3 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Agrawal H. C., Agrawal D. Proteolipid protein and DM-20 are synthesized by Schwann cells, present in myelin membrane, but they are not fatty acylated. Neurochem Res. 1991 Aug;16(8):855–858. doi: 10.1007/BF00965533. [DOI] [PubMed] [Google Scholar]
- Antoniou M., deBoer E., Habets G., Grosveld F. The human beta-globin gene contains multiple regulatory regions: identification of one promoter and two downstream enhancers. EMBO J. 1988 Feb;7(2):377–384. doi: 10.1002/j.1460-2075.1988.tb02824.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Aoyama A., Tamura T. A., Mikoshiba K. Regulation of brain-specific transcription of the mouse myelin basic protein gene: function of the NFI-binding site in the distal promoter. Biochem Biophys Res Commun. 1990 Mar 16;167(2):648–653. doi: 10.1016/0006-291x(90)92074-a. [DOI] [PubMed] [Google Scholar]
- Behringer R. R., Hammer R. E., Brinster R. L., Palmiter R. D., Townes T. M. Two 3' sequences direct adult erythroid-specific expression of human beta-globin genes in transgenic mice. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7056–7060. doi: 10.1073/pnas.84.20.7056. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bonnerot C., Grimber G., Briand P., Nicolas J. F. Patterns of expression of position-dependent integrated transgenes in mouse embryo. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6331–6335. doi: 10.1073/pnas.87.16.6331. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Braun P. E., Sandillon F., Edwards A., Matthieu J. M., Privat A. Immunocytochemical localization by electron microscopy of 2'3'-cyclic nucleotide 3'-phosphodiesterase in developing oligodendrocytes of normal and mutant brain. J Neurosci. 1988 Aug;8(8):3057–3066. doi: 10.1523/JNEUROSCI.08-08-03057.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brinster R. L., Allen J. M., Behringer R. R., Gelinas R. E., Palmiter R. D. Introns increase transcriptional efficiency in transgenic mice. Proc Natl Acad Sci U S A. 1988 Feb;85(3):836–840. doi: 10.1073/pnas.85.3.836. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen-Kiang S., Lavery D. J. Pulse labeling of heterogeneous nuclear RNA in isolated nuclei. Methods Enzymol. 1989;180:82–96. doi: 10.1016/0076-6879(89)80094-1. [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]
- Devine-Beach K., Lashgari M. S., Khalili K. Myelin basic protein gene transcription. Identification of proximal and distal cis-acting regulatory elements. J Biol Chem. 1990 Aug 15;265(23):13830–13835. [PubMed] [Google Scholar]
- Franklin G. C., Donovan M., Adam G. I., Holmgren L., Pfeifer-Ohlsson S., Ohlsson R. Expression of the human PDGF-B gene is regulated by both positively and negatively acting cell type-specific regulatory elements located in the first intron. EMBO J. 1991 Jun;10(6):1365–1373. doi: 10.1002/j.1460-2075.1991.tb07656.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Friedrich G., Soriano P. Promoter traps in embryonic stem cells: a genetic screen to identify and mutate developmental genes in mice. Genes Dev. 1991 Sep;5(9):1513–1523. doi: 10.1101/gad.5.9.1513. [DOI] [PubMed] [Google Scholar]
- Friedrich V. L., Jr, Centor D., Chambers J., Arnheiter H., Lazzarini R. A. Expression of exogenous glycoprotein genes in oligodendrocytes of transgenic mice. Ann N Y Acad Sci. 1990;605:262–269. doi: 10.1111/j.1749-6632.1990.tb42399.x. [DOI] [PubMed] [Google Scholar]
- Fritz R. B., Chou C. H. Epitopes of peptide 43-88 of guinea pig myelin basic protein: localization with monoclonal antibodies. J Immunol. 1983 May;130(5):2180–2182. [PubMed] [Google Scholar]
- Gillies S. D., Morrison S. L., Oi V. T., Tonegawa S. A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell. 1983 Jul;33(3):717–728. doi: 10.1016/0092-8674(83)90014-4. [DOI] [PubMed] [Google Scholar]
- Godfraind C., Friedrich V. L., Holmes K. V., Dubois-Dalcq M. In vivo analysis of glial cell phenotypes during a viral demyelinating disease in mice. J Cell Biol. 1989 Nov;109(5):2405–2416. doi: 10.1083/jcb.109.5.2405. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grosschedl R., Baltimore D. Cell-type specificity of immunoglobulin gene expression is regulated by at least three DNA sequence elements. Cell. 1985 Jul;41(3):885–897. doi: 10.1016/s0092-8674(85)80069-6. [DOI] [PubMed] [Google Scholar]
- Herman G. E., O'Brien W. E., Beaudet A. L. An E. coli beta-galactosidase cassette suitable for study of eukaryotic expression. Nucleic Acids Res. 1986 Sep 11;14(17):7130–7130. doi: 10.1093/nar/14.17.7130. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horrocks L. A. Composition of mouse brain myelin during development. J Neurochem. 1968 Jun;15(6):483–488. doi: 10.1111/j.1471-4159.1968.tb08945.x. [DOI] [PubMed] [Google Scholar]
- Johnson G. D., Davidson R. S., McNamee K. C., Russell G., Goodwin D., Holborow E. J. Fading of immunofluorescence during microscopy: a study of the phenomenon and its remedy. J Immunol Methods. 1982 Dec 17;55(2):231–242. doi: 10.1016/0022-1759(82)90035-7. [DOI] [PubMed] [Google Scholar]
- Katsuki M., Sato M., Kimura M., Yokoyama M., Kobayashi K., Nomura T. Conversion of normal behavior to shiverer by myelin basic protein antisense cDNA in transgenic mice. Science. 1988 Jul 29;241(4865):593–595. doi: 10.1126/science.2456614. [DOI] [PubMed] [Google Scholar]
- Kimura M., Sato M., Akatsuka A., Nozawa-Kimura S., Takahashi R., Yokoyama M., Nomura T., Katsuki M. Restoration of myelin formation by a single type of myelin basic protein in transgenic shiverer mice. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5661–5665. doi: 10.1073/pnas.86.14.5661. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987 Oct 26;15(20):8125–8148. doi: 10.1093/nar/15.20.8125. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lees M. B., Paxman S. Modification of the Lowry procedure for the analysis of proteolipid protein. Anal Biochem. 1972 May;47(1):184–192. doi: 10.1016/0003-2697(72)90291-6. [DOI] [PubMed] [Google Scholar]
- Lemke G. Unwrapping the genes of myelin. Neuron. 1988 Sep;1(7):535–543. doi: 10.1016/0896-6273(88)90103-1. [DOI] [PubMed] [Google Scholar]
- Magram J., Niederreither K., Costantini F. Beta-globin enhancers target expression of a heterologous gene to erythroid tissues of transgenic mice. Mol Cell Biol. 1989 Oct;9(10):4581–4584. doi: 10.1128/mcb.9.10.4581. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miura M., Tamura T., Aoyama A., Mikoshiba K. The promoter elements of the mouse myelin basic protein gene function efficiently in NG108-15 neuronal/glial cells. Gene. 1989 Jan 30;75(1):31–38. doi: 10.1016/0378-1119(89)90380-6. [DOI] [PubMed] [Google Scholar]
- Molineaux S. M., Engh H., de Ferra F., Hudson L., Lazzarini R. A. Recombination within the myelin basic protein gene created the dysmyelinating shiverer mouse mutation. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7542–7546. doi: 10.1073/pnas.83.19.7542. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Puckett C., Hudson L., Ono K., Friedrich V., Benecke J., Dubois-Dalcq M., Lazzarini R. A. Myelin-specific proteolipid protein is expressed in myelinating Schwann cells but is not incorporated into myelin sheaths. J Neurosci Res. 1987;18(4):511–518. doi: 10.1002/jnr.490180402. [DOI] [PubMed] [Google Scholar]
- Queen C., Baltimore D. Immunoglobulin gene transcription is activated by downstream sequence elements. Cell. 1983 Jul;33(3):741–748. doi: 10.1016/0092-8674(83)90016-8. [DOI] [PubMed] [Google Scholar]
- Readhead C., Popko B., Takahashi N., Shine H. D., Saavedra R. A., Sidman R. L., Hood L. Expression of a myelin basic protein gene in transgenic shiverer mice: correction of the dysmyelinating phenotype. Cell. 1987 Feb 27;48(4):703–712. doi: 10.1016/0092-8674(87)90248-0. [DOI] [PubMed] [Google Scholar]
- Roach A., Takahashi N., Pravtcheva D., Ruddle F., Hood L. Chromosomal mapping of mouse myelin basic protein gene and structure and transcription of the partially deleted gene in shiverer mutant mice. Cell. 1985 Aug;42(1):149–155. doi: 10.1016/s0092-8674(85)80110-0. [DOI] [PubMed] [Google Scholar]
- Russo A. F., Crenshaw E. B., 3rd, Lira S. A., Simmons D. M., Swanson L. W., Rosenfeld M. G. Neuronal expression of chimeric genes in transgenic mice. Neuron. 1988 Jun;1(4):311–320. doi: 10.1016/0896-6273(88)90079-7. [DOI] [PubMed] [Google Scholar]
- Sternberger N. H., Itoyama Y., Kies M. W., Webster H deF Immunocytochemical method to identify basic protein in myelin-forming oligodendrocytes of newborn rat C.N.S. J Neurocytol. 1978 Apr;7(2):251–263. doi: 10.1007/BF01217922. [DOI] [PubMed] [Google Scholar]
- Sturrock R. R. Myelination of the mouse corpus callosum. Neuropathol Appl Neurobiol. 1980 Nov-Dec;6(6):415–420. doi: 10.1111/j.1365-2990.1980.tb00219.x. [DOI] [PubMed] [Google Scholar]
- Tamura T., Aoyama A., Inoue T., Miura M., Okano H., Mikoshiba K. Tissue-specific in vitro transcription from the mouse myelin basic protein promoter. Mol Cell Biol. 1989 Jul;9(7):3122–3126. doi: 10.1128/mcb.9.7.3122. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tamura T., Mikoshiba K. Demonstration of a transcription element in vitro between the capping site and translation initiation site of the mouse myelin basic protein gene. FEBS Lett. 1991 Mar 11;280(1):75–78. doi: 10.1016/0014-5793(91)80207-j. [DOI] [PubMed] [Google Scholar]
- Tamura T., Sumita K., Hirose S., Mikoshiba K. Core promoter of the mouse myelin basic protein gene governs brain-specific transcription in vitro. EMBO J. 1990 Oct;9(10):3101–3108. doi: 10.1002/j.1460-2075.1990.tb07507.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Timmons T. M., Dunbar B. S. Protein blotting and immunodetection. Methods Enzymol. 1990;182:679–688. doi: 10.1016/0076-6879(90)82053-5. [DOI] [PubMed] [Google Scholar]
- Turnley A. M., Morahan G., Okano H., Bernard O., Mikoshiba K., Allison J., Bartlett P. F., Miller J. F. Dysmyelination in transgenic mice resulting from expression of class I histocompatibility molecules in oligodendrocytes. Nature. 1991 Oct 10;353(6344):566–569. doi: 10.1038/353566a0. [DOI] [PubMed] [Google Scholar]
- Whitaker J. N. The protein antigens of peripheral nerve myelin. Ann Neurol. 1981;9 (Suppl):56–64. doi: 10.1002/ana.410090710. [DOI] [PubMed] [Google Scholar]
- Wiktorowicz M., Roach A. Regulation of myelin basic protein gene transcription in normal and shiverer mutant mice. Dev Neurosci. 1991;13(3):143–150. doi: 10.1159/000112152. [DOI] [PubMed] [Google Scholar]
- Yoshioka T., Feigenbaum L., Jay G. Transgenic mouse model for central nervous system demyelination. Mol Cell Biol. 1991 Nov;11(11):5479–5486. doi: 10.1128/mcb.11.11.5479. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Ferra F., Engh H., Hudson L., Kamholz J., Puckett C., Molineaux S., Lazzarini R. A. Alternative splicing accounts for the four forms of myelin basic protein. Cell. 1985 Dec;43(3 Pt 2):721–727. doi: 10.1016/0092-8674(85)90245-4. [DOI] [PubMed] [Google Scholar]