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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Journal of neurochemistry 83 (2002), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Phosphatidylinositol-3 kinase (PI3K) is a family of enzymes that phosphorylates the D3 position of phosphoinositides in membranes which can then act as a second messenger and affect many essential cellular processes such as survival, proliferation and differentiation. Class IA PI3K is composed of two subunits: a regulatory subunit, p85, and a catalytic subunit, p110. The p85 subunit is composed of several adapter domains which, upon interaction with the appropriate molecules, transmit the signal to activate p110. We have used the spontaneously immortalized oligodendrocyte cell line, CG4, to examine the role of PI3K in maturation of the oligodendrocyte. We show that overexpression of the p85 subunit enhances expression of myelin basic protein (MBP) upon differentiation of CG4 cells and primary oligodendrocytes. In experiments in CG4 cells, neither cotransfection with the tumor suppressor PTEN, which dephosphorylates the D3 position of phosphoinositides, nor inhibition of PI3K activity with wortmannin mimics this effect. Further, we have shown that this effect is dependent on the coexpression of the two SH2 domains within p85. Thus, the p85-mediated enhancement of MBP promoter activity in oligodendrocytes appears to be independent of PI3K activity and dependent on the adapter functions of the p85 subunit's SH2 domains.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 112 (1982), S. 327-338 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The binding, internalization, intracellular translocation, and degradation of epidermal growth factor (EGF) were studied in mouse Swiss/3T3 fibroblasts under two different physiological conditions at 37°C. In serum-containing medium the maximal level of cell-bound EGF was maintained for at least 8 h without appreciable degradation in contrast to serum-free conditions. These phenomena were correlated with a difference in the intracellular site to which the receptor-bound EGF was delivered as studied using Percoll density gradients. In serum-containing medium the majority of cell-bound EGF was initially taken up into a Golgi-like vesicle of density 1.046, corresponding to the marker galactosyl transferase, and then delivered to a population of vesicles with similar density as lysosomes (ϱ =1.068-1.110). A portion of the EGF became degraded and was released from the cell into the medium while the remainder stayed with the cells, intact, for a long period of time. In serum-free medium, EGF became associated with a heterogeneous population of vesicles with a mean density of 1.050 which do not correspond to any of the marker enzymes for subcellular organelles for which we have tested (Golgi, endoplasmic reticulum, plasma membrane, lysosomes). It is then transferred to lysosome-like vesicles (ϱ = 1.068-1.110). We therefore propose that EGF is processed through two separate endocytotic routes which are regulated by the cell depending upon its physiological state.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The internalization process following cell surface receptor binding by epidermal growth factor (EGF) was studied. It was found that EGF is taken up into a dense, membranous organelle. This organelle is deficient in lysosomal enzyme activity and is biochemically dissimilar to the major lysosomal fraction. The uptake of EGF by this organelle was demonstrated in three different cell types representing three different species. Each of these cell types is highly responsive to the mitogenic action of EGF. These results indicate that EGF is endocytosed and delivered to a dense, possibly nonlysosomal, organelle which is ubiquitous in fibroblasts. Furthermore, we demonstrate a close, positive correlation between EGF uptake into this fraction and the ability of cells to respond to the mitogen. A negative relationship between uptake into the subcellular fraction containing lysosomal enzymes and EGF-stimulated DNA synthesis was observed. Using numerous incubation conditions no exceptions to the correlation between EFG uptake into the lysosomal enzymedeficient fraction and EGF-induced DNA synthesis were observed. These results suggest a role for this dense organelle in the production of a mitogenic signal.
    Additional Material: 13 Ill.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 49 (1992), S. 349-356 
    ISSN: 0730-2312
    Keywords: Transcription factors ; Sp1 ; Ap1 ; CREB ; DNase I footprinting ; Oligonucleotide competition ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Nuclear factors from HeLa cells were isolated by elution of DNA-cellulose bound proteins with a double stranded synthetic oligonucleotide corresponding to the region from -34 to -79 of the human transferrin receptor (TR) gene promoter. The eluted proteins were further purified and separated from the oligonucleotide by ion exchange chromatography. Proteins within the resulting fraction bound with specificity to the TR promoter. Retardation gel analysis and competition with specific double-stranded oligonucleotides show that multiple factors present in this fraction compete for binding within the same region of the TR promoter. Footprinting experiments demonstrate that these factors contact a GC-rich element that is within the region that is required for enhanced expression of the gene in proliferating cells. One of the factors protects an extended DNA sequence but, still contacts the GC-rich element.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 20 (1982), S. 41-50 
    ISSN: 0730-2312
    Keywords: hormone receptors ; Golgi ; lysosomes ; Percoll gradient ; endocytosis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The intracellular translocation and processing of epidermal growth factor (EOF) in 3T3 cells has been studied utilizing Percoll density gradients. EGF is internalized and rapidly becomes associated with two types of intracellular compartments. The extent to which EGF is delivered to these two compartments is apparently regulated depending upon the cell's physiological condition. In growth medium, an increased proportion of EGF is taken up into a Golgi-like element. Uptake through this pathway correlates with a decrease in degradation of the ligand. In the absence of scrum and amino acids, an increased proportion of EGF is taken up into a component which has a density of 1.05. Uptake through this pathway correlates with increased degradation of the ligand. The ligand taken up through both pathways is transferred to dense vesicles which comigrate with lysosomes. In the presence of growth medium, however, dense vesicles containing EGF can be shown to be lysosomal enzyme-deficient upon further fractionation. In addition, in the presence of serum, a portion of the internalized EGF is apparently released from the cells, intact, and then re-bound. The processes described may be important in the production of a mitogenic response and the ability of cells to self-regulate their responsiveness to the growth factor.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Supramolecular Structure and Cellular Biochemistry 16 (1981), S. 105-113 
    ISSN: 0275-3723
    Keywords: hormone receptors ; diphtheria toxin ; lysosomes ; hybrid proteins ; Chemistry ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Recently we have isolated six variants of Swiss/3T3 mouse fibroblasts that are resistant to the cytotoxic insulin-diphtheria toxin A fragment. All of the variants proved to have greatly reduced or no insulin binding capacity, and several variants showed altered morphologies and growth characteristics. We now report on the further characterization of one of these variants, CI-3. which displays a massive accumulation of membranous vesicles in its cytoplasm. By electron microscopy these vesicles resemble lysosomes. They also appear to fluorcsce bright orange after treatment of viable cells with acridine orange. However, the specific activity of several lysosomal enzymes is depressed in CI-3. Additionally, there is an apparent shift in the density of vesicles containing lysosomal enzymes in this variant. These alterations may be directly related to CI-3′s resistance to the cytotoxic insulin and have some important bearings on the mechanism of insulin action.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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