ras isoprenylation is required for ras-induced but not for NGF-induced neuronal differentiation of PC12 cells

M S Qiu; A F Pitts; T R Winters; S H Green

doi:10.1083/jcb.115.3.795

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ras isoprenylation is required for ras-induced but not for NGF-induced neuronal differentiation of PC12 cells

Journal article

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Peer reviewed

ras isoprenylation is required for ras-induced but not for NGF-induced neuronal differentiation of PC12 cells

M S Qiu, A F Pitts, T R Winters and S H Green

The Journal of cell biology, Vol.115(3), pp.795-808

11/1991

DOI: 10.1083/jcb.115.3.795

PMCID: PMC2289192

PMID: 1918164

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Abstract

We have used compactin, an inhibitor of mevalonate biosynthesis, to block p21ras posttranslational modification and membrane association in PC12 cells. Previous studies have demonstrated a requirement for isoprenylation for mitogenic effects of activated p21ras in mammalian cells and for function of RAS gene products in yeast. Immunoprecipitation of [35S]methionine-labeled p21ras from PC12 cell homogenates confirmed that the processed p21ras species is missing from compactin-treated PC12 cells. Immunoprecipitation from particulate and cytosolic fractions of PC12 cells confirmed that compactin blocks p21ras membrane association: p21ras is confined to the cytosol fraction. Induction of neuronal differentiation and ornithine decarboxylase (ODCase) transcription by oncogenic p21N-ras does not occur in compactin-treated cells indicating that activity of oncogenic p21N-ras expressed in PC12 cells is abolished by compactin treatment. Thus, p21ras isoprenylation or association with the membrane appears to be required for early responses and neuronal differentiation attributable to p21ras activation. In contrast, blockade of p21ras isoprenylation and membrane association by compactin treatment did not significantly reduce PC12 cell responses to NGF. Responses examined included rapid phosphorylation of tyrosine hydroxylase, rapid induction of ODCase expression, survival in serum-free medium and neuronal differentiation. Compactin blocked growth factor-induced rapid changes in cell surface morphology but did so whether this response was induced by NGF or by EGF. These results indicate that functional p21ras is not necessary for responses to NGF which in turn implies that if a ras-dependent NGF signal transduction pathway exists, as has been previously suggested, at least one additional ras-independent pathway must also be present.

Animals

Cell Differentiation - drug effects

Cytosol - physiology

Dexamethasone - pharmacology

DNA Probes

Epidermal Growth Factor - pharmacology

Gene Expression - drug effects

Genes, ras

Hydroxymethylglutaryl-CoA Reductase Inhibitors

Lovastatin - analogs & derivatives

Lovastatin - pharmacology

Nerve Growth Factors - pharmacology

Neurons - cytology

Neurons - drug effects

Ornithine Decarboxylase - genetics

PC12 Cells

Phosphoproteins - biosynthesis

Phosphoproteins - isolation & purification

Polyisoprenyl Phosphates - metabolism

Protein Processing, Post-Translational - drug effects

Proto-Oncogene Proteins p21(ras) - genetics

RNA, Neoplasm - genetics

RNA, Neoplasm - isolation & purification

Signal Transduction - drug effects

Details

Title: Subtitle: ras isoprenylation is required for ras-induced but not for NGF-induced neuronal differentiation of PC12 cells
Creators: M S Qiu - Department of Biology, University of Iowa, Iowa City 52242
A F Pitts
T R Winters
S H Green
Resource Type: Journal article
Publication Details: The Journal of cell biology, Vol.115(3), pp.795-808
DOI: 10.1083/jcb.115.3.795
PMID: 1918164
PMCID: PMC2289192
ISSN: 0021-9525
eISSN: 1540-8140
Grant note: MH14629 / NIMH NIH HHS NS26538 / NINDS NIH HHS
Language: English
Date published: 11/1991
Academic Unit: Biology; Otolaryngology
Record Identifier: 9984217424502771

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