Mitogenic hormone-induced intracellular message: assay and partial characterization of an activator of DNA replication induced by epidermal growth factor.

Das, Manjusri

doi:10.1073/pnas.77.1.112

Cited by 49 publications

(10 citation statements)

References 19 publications

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“…As shown by Ohkuma & Poole (1978) this process results in an increase of pH to values outside the optimal range of the acid hydrolases and may thus interfere with lysosomal function. It has been proposed that continuous degradation or limited processing of ligand-receptor complexes by lysosomal enzymes is required to stimulate cell growth, one hypothesis being that this results in the production of a necessary intracellular messenger (Das, 1980;King et al, 1980). It is also possible that general lysosomal activity is essential for the lymphocyte to reprocess its constituents during activation and that inhibition of the acid hydrolases prevents this.…”

Section: Discussionmentioning

confidence: 99%

“…Indeed, chloroquine and other lysosomotropic amines have been shown to inhibit the mitogenesis of fibroblasts induced by epidermal growth factor (King etal., 1980). It is possible that degradation of the internalized ligand-receptor complex by the acid hydrolases in the lysosomes may be an essential step in the transduction mechanism of the mitogenic response to a ligand (Das, 1980). If this is so, then other simpler amines should be equally effective in inhibiting lymphocyte activation.…”

Section: Intoductionmentioning

confidence: 99%

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A comparison of dimaprit, nordimaprit, methylamine and chloroquine as inhibitors of mitogen‐induced lymphocyte activation

Dale

Ladd

1984

British J Pharmacology

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Methylamine and chloroquine both ‘lysosomotropic’ agents (i.e. agents which sequester in lysosomes) caused a dose‐related inhibition of mitogen‐induced lymphocyte activation in the concentrations which have previously been shown to increase the pH of lysosomes. The dose‐response curves of inhibition of mitogen‐induced lymphocyte activation for chloroquine and methylamine are very steep and are similar to the dose‐response curves obtained with dimaprit and nordimaprit, but very different from the flat dose‐response curves previously described for histamine. Approximate IC50 values were methylamine 6.4 mm, dimaprit 0.13 mm, nordimaprit 0.03 mm and chloroquine 18 μm. It is suggested that the mechanism of action of methylamine and chloroquine may be related to their lysosomotropic action and consequent interference with ligand‐receptor processing, and that dimaprit and nordimaprit but not histamine may act by a similar mechanism.

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Section: Discussionmentioning

confidence: 99%

Section: Intoductionmentioning

confidence: 99%

A comparison of dimaprit, nordimaprit, methylamine and chloroquine as inhibitors of mitogen‐induced lymphocyte activation

Dale

Ladd

1984

British J Pharmacology

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“…Other studies have focused on growth factor-stimulated initiation of DNA synthesis by using nuclei from Xenopus laevis spleen cells (17) as a native DNA template. However, since Xenopus chromosomes may not have specific origins of replication (18), it is not clear whether stimulation ofXenopus nuclear replication by subcellular fractions from mitogenstimulated mammalian cells reflects critical cell-cycle control mechanisms operative in mammalian cells.…”

Section: Discussionmentioning

confidence: 99%

Growth factor-dependent initiation of DNA replication in nuclei isolated from an interleukin 3-dependent murine myeloid cell line

1990

J. Clin. Invest.

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To study the proliferative response of hematopoietic cells to growth factors at the molecular level, we developed a cell-free system for growth factor-dependent initiation of genomic DNA replication. Nuclei were isolated from the IL-3-dependent cell line NFS/N1-H7 after a 10-h period of IL-3 deprivation. Cytosolic and membrane-containing subcellular fractions were prepared from proliferating NFS/N1-H7 cells. Nuclei from the nonproliferating cells (±IL-3) showed essentially no incorporation of VH]thymidine during a 16-h incubation with a mixture of unlabeled GTP, ATP, UTP, CITP, dGTP, dATP, dCTP, and VHldTTP. When the combination of IL-3, a cytosolic fraction, and a membrane-containing fraction from proliferating cells was added to nuclei from nonproliferating cells, a burst of VIHjthymidine incorporation into DNA began after a 12-h lag period, attained a maximal rate at 16 h, and reached a level of 860 pmol thymidine/10' nuclei at 24 h (corresponding to replication of -56% total mouse genomic DNA). This DNA synthesis was inhibited 90% by the specific DNA polymerase a inhibitor aphidicolin. Deletion of a single cellular component or IL-3 from the system resulted in a marked reduction of DNA replication (-membrane, 80±4%; -cytosol, 90%±4%; -IL-3, 74±7% inhibition). This model requires a growth factor (IL-3), a sedimentable cell fraction containing its receptor and possibly additional membrane-associated components, and a cytosolic fraction. It appears to recapitulate the molecular events required for progression from early GI to S phase of the cell cycle induced by IL-3 binding to its receptor. (J. Clin. Invest. 1990. 85:300-304.) cell-free system * DNA replication -growth factor * IL-3

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“…For example, we have shown that spontaneously proliferating lymphoblastoid cells, PHA-stimulated human peripheral blood lymphocytes (PBL), and interleukin 2 (IL-2)-activated T cell blasts contain an extractable cytoplasmic protein called ADR Receivedfor publication 26 November 1985 and in revisedform 13 February 1986.…”

Section: Introductionmentioning

confidence: 99%

“…The assay used was a modification of the procedure of Das (13 15-min incubatiop at 0°C, the cultures were harvested as described above. The evidence that this assay measures DNA replication has been summarized (14), and includes the absolute requirement for ATP and the presence of replicative forks and "eyes."…”

Section: Introductionmentioning

confidence: 99%

Impaired nuclear responsiveness to cytoplasmic signals in lymphocytes from elderly humans with depressed proliferative responses.

Gutowski¹,

Innes²,

Weksler³

et al. 1986

J. Clin. Invest.

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Lymphocytes from many elderly individuals exhibit depressed proliferative responses to the plant phytohemagglutinin (PHA). We have previously reported that this proliferative defect is not due to a failure to generate a cytoplasmic activator of DNA replication (ADR). In the present study, we tested the DNA synthetic response of nuclei derived from aged cells to an exogenous source of ADR. We found that nuclei from aged lymphocytes exhibiting low PHA responses were impaired in their ability to synthesize DNA in response to ADR, compared with nuclei from younger adult donors. In contrast, those aged cells maintaining intact PHA responses provided nuclei that were also unimpaired in their response to ADR. The relationship between PHA responsiveness of the intact cells and ADR responsiveness of nuclei derived from these was a linear one. These results suggest that the depressed cellular reactivity of aged lymphocytes to PHA (when seen) may be due to a failure of these nuclei to respond to cytoplasmic stimulatory signals induced by the mitogen.

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Mitogenic hormone-induced intracellular message: assay and partial characterization of an activator of DNA replication induced by epidermal growth factor.

Abstract: This paper explores the pathway from nuclear quiescence to mitogenesis. It describes an in vitro assay for an activator of DNA replication induced by epidermal growth factor (EGF)

Cited by 49 publications

References 19 publications

A comparison of dimaprit, nordimaprit, methylamine and chloroquine as inhibitors of mitogen‐induced lymphocyte activation

A comparison of dimaprit, nordimaprit, methylamine and chloroquine as inhibitors of mitogen‐induced lymphocyte activation

Growth factor-dependent initiation of DNA replication in nuclei isolated from an interleukin 3-dependent murine myeloid cell line

Impaired nuclear responsiveness to cytoplasmic signals in lymphocytes from elderly humans with depressed proliferative responses.

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