The treatment of density-arrested BALB/c-3T3 cells with electrophoretically homogenous or highly purified preparations of the platelet-derived growth factor (PDGF) stimulated the rapid and selective accumulation of several species of abundant mRNA identified by cell-free translation. These translatable mRNAs appeared long before entry into the S phase. Less PDGF was required for selective mRNA accumulation than for PDGF-modulated DNA synthesis. The translatable mRNAs also accumulated after addition of the epidermal growth factor but not after addition of insulin or platelet-poor plasma. Their selective accumulation was blocked by addition of actinoimycin D. Three classes of PDGF-modulated mRNAs were defined. An early (primary) RNA appeared within 30 to 60 min of PDGF addition; its accumulation was not blocked by cycloheximide. Another early mRNA also appeared within 60 min, but treatment with both PDGF and cycloheximide was required for optimal accumulation. A third class, secondary RNAs, began to accumulate later at 90 to 120 min; the appearance of this class was inhibited by cycloheximide. One- and two-dimensional gel electrophoresis of translation products demonstrated that a spontaneously transformed BALB/c-3T3 (ST2-3T3) cell line, which does not require PDGF or EGF for growth, constitutively accumulated the secondary growth factor-regulated mRNAs. The accumulation of these translatable mRNAs may be required for PDGF-modulated DNA synthesis. We have recently utilized a cDNA probe of a PDGF-regulated lysosomal protein (termed MEP) to directly quantify a PDGF-modulated mRNA. PDGF began to stimulate MEP mRNA accumulation 240 min after addition in a dose dependent fashion. Other growth factors, including EGF, IGF-1, insulin, and platelet-poor plasma did not have this effect. The PDGF modulated accumulation of MEP mRNA was inhibitable by cycloheximide demonstrating that PDGF modulated protein synthesis is required. A spontaneously transformed variant of BALB/c-3T3 cells which does not require PDGF for growth accumulated MEP mRNA in a constitutive fashion. Thus the MEP transcript is an example of a PDGF-modulated secondary RNA. (J)

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA034162-05
Application #
3171942
Study Section
Molecular Biology Study Section (MBY)
Project Start
1982-07-01
Project End
1987-06-30
Budget Start
1986-03-01
Budget End
1987-06-30
Support Year
5
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Children's Hospital of Philadelphia
Department
Type
DUNS #
073757627
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Frick, K K; Scher, C D (1990) Platelet-derived growth factor-stimulated c-myc RNA accumulation in MG-63 human osteosarcoma cells is independent of both protein kinase A and protein kinase C. Mol Cell Biol 10:184-92
Ruggeri, B A; Klurfeld, D M; Kritchevsky, D et al. (1990) Steady-state mRNA expression for growth factors in DMBA-induced rat mammary tumors. Cancer Lett 55:89-93
Grundy, P; Bishayee, S; Disa, S et al. (1989) Modulation of platelet-derived growth factor receptor function in BP3T3, a chemically transformed BALB/c-3T3 cell line. Cancer Res 49:3581-6
Bishayee, S; Majumdar, S; Scher, C D et al. (1988) Characterization of a novel anti-peptide antibody that recognizes a specific conformation of the platelet-derived growth factor receptor. Mol Cell Biol 8:3696-702
Scher, C D (1988) The platelet derived growth factor. Mead Johnson Symp Perinat Dev Med :28-32
Frick, K K; Womer, R B; Scher, C D (1988) Platelet-derived growth factor-induced c-myc RNA expression. Analysis of an inducible pathway independent of protein kinase C. J Biol Chem 263:2948-52
Whipple, A P; Furlanetto, R W; Scher, C D (1987) Conditional responsiveness of a chemically transformed cell line to growth factor stimulated DNA synthesis. J Cell Physiol 130:182-90
Womer, R B; Frick, K; Mitchell, C D et al. (1987) PDGF induces c-myc mRNA expression in MG-63 human osteosarcoma cells but does not stimulate cell replication. J Cell Physiol 132:65-72
Scher, C D; Engle, L J; Eberenz, W M et al. (1986) Dissociation of cellular transformation from platelet-derived growth factor independence. J Cell Physiol 126:333-40
Bishayee, S; Ross, A H; Womer, R et al. (1986) Purified human platelet-derived growth factor receptor has ligand-stimulated tyrosine kinase activity. Proc Natl Acad Sci U S A 83:6756-60

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