In this application, we propose to test two new hypotheses concerning the MBP gene. These hypotheses are based upon data derived from the identification of two classes of unusual myelin basic protein (MBP)-related transcripts isolated from screens of mouse brain and human fetal spinal cord cDNA libraries. The first hypothesis is that the structure of the MBP gene is presently incomplete and contains at least one more exon upstream of what is currently believed to be exon 1 and an additional promoter. The second hypothesis is that portions of the MBP gene (including parts of at least one exon and one intron) are transcribed into mRNAs encoding proteins other than the MBP variants characterized thus far. The location of any additional exon sequences will be determined by conventional procedures of mapping and sequencing genomic clones using an MBP cDNA that contains sequence corresponding to the additional exon region. Sites of transcription initiation at this new upstream exon will be determined by S-1 nuclease and primer extension analysis. Experiments will be performed to determine if initiation of transcription at this start site and the one previously identified in the MBP gene varies during development or in different regions of the CNS. Experiments will also be performed to determine if all MBP variant forms are produced when transcription begins at this new transcription site. Full length cDNAs corresponding to the second class of MBP-gene related transcripts will be isolated, sequenced, and characterized. The structural relationship between this second class of transcripts and the MBP gene will be determined. The intron/exon arrangement of the gene coding for these MBP-related cDNAs will be elucidated and it will be determined if all the exons of this gene are encoded by genomic segments completely included within the MBP gene or if some exons lie outside the MBP gene. To determine if these transcripts are expressed in tissues other than brain or in cell types other than oligodendrocytes, Northern blot analyses of mRNA from a variety of tissues and cultured cells will be performed. The developmental appearance of the transcripts in the brains of normal mice and dysmyelinating mutants will be compared with the expression of the MBP mRNAs. In situ hybridization studies will be performed to determine the cellular and regional localization of the transcripts.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS023322-05
Application #
3406636
Study Section
Neurology C Study Section (NEUC)
Project Start
1985-07-01
Project End
1991-02-28
Budget Start
1990-03-01
Budget End
1991-02-28
Support Year
5
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Type
Other Domestic Higher Education
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Bongarzone, E R; Foster, L M; Byravan, S et al. (1997) Temperature-dependent regulation of PLP/DM20 and CNP gene expression in two conditionally-immortalized jimpy oligodendrocyte cell lines. Neurochem Res 22:363-72
Pribyl, T M; Campagnoni, C W; Kampf, K et al. (1996) Expression of the myelin proteolipid protein gene in the human fetal thymus. J Neuroimmunol 67:125-30
Pribyl, T M; Campagnoni, C W; Kampf, K et al. (1996) Expression of the myelin basic protein gene locus in neurons and oligodendrocytes in the human fetal central nervous system. J Comp Neurol 374:342-53
Landry, C F; Ellison, J A; Pribyl, T M et al. (1996) Myelin basic protein gene expression in neurons: developmental and regional changes in protein targeting within neuronal nuclei, cell bodies, and processes. J Neurosci 16:2452-62
Pribyl, T M; Campagnoni, C; Kampf, K et al. (1996) The major myelin protein genes are expressed in the human thymus. J Neurosci Res 45:812-9
Jacobs, E C; Arnold, A P; Campagnoni, A T (1996) Zebra finch estrogen receptor cDNA: cloning and mRNA expression. J Steroid Biochem Mol Biol 59:135-45
Foster, L M; Landry, C; Phan, T et al. (1995) Conditionally immortalized oligodendrocyte cell lines migrate to different brain regions and elaborate 'myelin-like' membranes after transplantation into neonatal shiverer mouse brains. Dev Neurosci 17:160-70
Ueno, S; Foster, L; Hifumi, G T et al. (1995) The simian virus 40 large T antigen does not inhibit translation of the 14-kDa myelin basic protein mRNA in reticulocyte lysates or in transfected cells. J Neurochem 64:928-31
Kashima, T; Vinters, H V; Campagnoni, A T (1995) Unexpected expression of intermediate filament protein genes in human oligodendroglioma cell lines. J Neuropathol Exp Neurol 54:23-31
Byravan, S; Campagnoni, A T (1994) Serum factors and hydrocortisone influence the synthesis of myelin basic proteins in mouse brain primary cultures. Int J Dev Neurosci 12:343-51

Showing the most recent 10 out of 43 publications