Expression of val-12 mutant RAS P21 in an IL-3-dependent murine myeloid cell line is associated with loss of serum-dependence and increases in membrane PIP2-specific phospholipase C activity

Maria Teresa Rizzo, H. Boswell, Denis English, Theodore G. Gabig

Research output: Contribution to journalArticle

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Abstract

We previously showed that the proliferative response of a serum- and interleukin-3 (IL-3)-dependent murine myeloid cell line, NFS/N1-H7, was partially inhibited by pertussis toxin as a result of toxin-induced increased adenylate cyclase activity. In the present studies, we examined the role of the phosphoinositide cycle in the proliferative response of these cells and demonstrated that there was no change in PIP (phosphatidyl-inositol bisphosphate)-specific phospholipase C activity in response to IL-3 alone. However, serum caused a pertussis toxin-insensitive increase in PIP2-specific phospholipase C activity as reflected by decreased cellular levels of 32P-labelled PIP2. Proliferation of a subline selected from val-12-mutant H-ras-transfected NFS-H7 cells, clone E5, was insensitive to pertussis toxin, occurred in the absence of serum but remained serum-stimulatable and absolutely dependent on IL-3. This val-12 mutant ras-expressing cell line showed an increase in 32P-labelled PIP (phosphatidylinositol phosphate) in response to serum whereas the parent cell line did not. Membrane fractions from 32P-labelled ras-transfected cells displayed higher GTPγS-, GTP-, or F- -stimulated PIP2-specific phospholipase C activity compared to membranes from the parent cell line. Thus serum-dependence and adenylate cyclase-mediated pertussis toxin-sensitivity of the parent cell line was bypassed by val-12 mutant ras p21, possibly as a result of increased PIP2-specific phospholipase C activity.

Original languageEnglish
Pages (from-to)311-319
Number of pages9
JournalCellular Signalling
Volume3
Issue number4
DOIs
StatePublished - 1991

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Interleukin-3
Type C Phospholipases
Myeloid Cells
Pertussis Toxin
Cell Line
Membranes
Serum
Phosphatidylinositols
Adenylyl Cyclases
Proto-Oncogene Proteins p21(ras)
Phosphatidylinositol Phosphates
Guanosine Triphosphate
Clone Cells

Keywords

  • pertussis toxin
  • Phosphoinositide cycle
  • phospholipase C
  • ras p 21

ASJC Scopus subject areas

  • Cell Biology

Cite this

Expression of val-12 mutant RAS P21 in an IL-3-dependent murine myeloid cell line is associated with loss of serum-dependence and increases in membrane PIP2-specific phospholipase C activity. / Rizzo, Maria Teresa; Boswell, H.; English, Denis; Gabig, Theodore G.

In: Cellular Signalling, Vol. 3, No. 4, 1991, p. 311-319.

Research output: Contribution to journalArticle

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