Differential regulation of fibroblast growth factor (FGF) receptor-1 mRNA and protein by two molecular forms of basic FGF

Modulation of FGFR-1 mRNA stability

Agnes Estival, Veronique Monzat, Karine Miquel, François Gaubert, Etienne Hollande, Murray Korc, Nicole Vaysse, François Clemente

Research output: Contribution to journalArticle

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Abstract

To evaluate possible functional differences between basic fibroblast growth factor (FGF) 2 isoforms we analyzed the effects of the 18-kDa FGF-2 which mainly localizes in the cytosol and that of the nuclear-targeted 22.5-kDa form on FGF receptors (FGFR) expression. These peptides were expressed at low amounts through a retroviral-infection system. Point mutated FGF-2 cDNAs under the control of the β-actin promoter were used to infect a pancreatic cell line (AR4-2J) which does not produce FGF-2. Saturation and competition binding studies with 125I-FGF-2 revealed a 3-fold increase in both high and low affinity receptors in cells expressing the 22.5-kDa form and a 2-fold increase only in the high affinity receptors in cells producing the 18-kDa form. Kd values and molecular weights of the high affinity receptors were unaffected. Increasing cell densities or cell treatment with exogenous FGF-2 resulted in FGFR down-regulation as in control cells. Neutralizing antiFGF-2 antibodies and suramin did not affect receptor density in control and in cells producing the 22.5-kDa form but further increased by 60 and 80%, respectively, the receptor level in cells synthesizing the 18-kDa form. These data suggest the involvement of the intracellular stored FGF-2 in FGFR up-regulation. Although all cells expressed FGFR-1, -2, and -3 mRNA only the FGFR-1 transcript was found increased, 6-fold in 22.5-kDa expressing cells and 3-fold in cell producing the shortest secreted isoform. The increase in FGFR-1 mRNA levels in the 22.5-kDa expressing cells was due to enhanced stability of the transcript. Confocal microscopy detected the presence of FGFR-1 at the cell surface whereas secretory isoforms of the receptor were not observed. Reverse transcriptase-polymerase chain reaction did not reveal significant differences in the expression of FGFR-1 variants. In the 22.5-kDa expressing cells exogenous FGF-2 evoked a stronger translocation of the calcium-phospholipid-dependent PEC. These results indicate that the transfected FGF-2 isoforms up-regulated FGFR-1 mRNA and protein. The 22.5-kDa form acted by increasing FGFR-1 mRNA stability enhancing cell responses to exogenous FGF-2.

Original languageEnglish (US)
Pages (from-to)5663-5670
Number of pages8
JournalJournal of Biological Chemistry
Volume271
Issue number10
StatePublished - Mar 8 1996
Externally publishedYes

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Receptor, Fibroblast Growth Factor, Type 1
Fibroblast Growth Factor 1
Fibroblast Growth Factor Receptors
RNA Stability
Fibroblast Growth Factor 2
Modulation
Messenger RNA
Proteins
Protein Isoforms
Cells
Suramin
Confocal microscopy
Polymerase chain reaction
RNA-Directed DNA Polymerase
Actins
Reverse Transcriptase Polymerase Chain Reaction
Phospholipids
Confocal Microscopy

ASJC Scopus subject areas

  • Biochemistry

Cite this

Differential regulation of fibroblast growth factor (FGF) receptor-1 mRNA and protein by two molecular forms of basic FGF : Modulation of FGFR-1 mRNA stability. / Estival, Agnes; Monzat, Veronique; Miquel, Karine; Gaubert, François; Hollande, Etienne; Korc, Murray; Vaysse, Nicole; Clemente, François.

In: Journal of Biological Chemistry, Vol. 271, No. 10, 08.03.1996, p. 5663-5670.

Research output: Contribution to journalArticle

Estival, A, Monzat, V, Miquel, K, Gaubert, F, Hollande, E, Korc, M, Vaysse, N & Clemente, F 1996, 'Differential regulation of fibroblast growth factor (FGF) receptor-1 mRNA and protein by two molecular forms of basic FGF: Modulation of FGFR-1 mRNA stability', Journal of Biological Chemistry, vol. 271, no. 10, pp. 5663-5670.
Estival, Agnes ; Monzat, Veronique ; Miquel, Karine ; Gaubert, François ; Hollande, Etienne ; Korc, Murray ; Vaysse, Nicole ; Clemente, François. / Differential regulation of fibroblast growth factor (FGF) receptor-1 mRNA and protein by two molecular forms of basic FGF : Modulation of FGFR-1 mRNA stability. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 10. pp. 5663-5670.
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abstract = "To evaluate possible functional differences between basic fibroblast growth factor (FGF) 2 isoforms we analyzed the effects of the 18-kDa FGF-2 which mainly localizes in the cytosol and that of the nuclear-targeted 22.5-kDa form on FGF receptors (FGFR) expression. These peptides were expressed at low amounts through a retroviral-infection system. Point mutated FGF-2 cDNAs under the control of the β-actin promoter were used to infect a pancreatic cell line (AR4-2J) which does not produce FGF-2. Saturation and competition binding studies with 125I-FGF-2 revealed a 3-fold increase in both high and low affinity receptors in cells expressing the 22.5-kDa form and a 2-fold increase only in the high affinity receptors in cells producing the 18-kDa form. Kd values and molecular weights of the high affinity receptors were unaffected. Increasing cell densities or cell treatment with exogenous FGF-2 resulted in FGFR down-regulation as in control cells. Neutralizing antiFGF-2 antibodies and suramin did not affect receptor density in control and in cells producing the 22.5-kDa form but further increased by 60 and 80{\%}, respectively, the receptor level in cells synthesizing the 18-kDa form. These data suggest the involvement of the intracellular stored FGF-2 in FGFR up-regulation. Although all cells expressed FGFR-1, -2, and -3 mRNA only the FGFR-1 transcript was found increased, 6-fold in 22.5-kDa expressing cells and 3-fold in cell producing the shortest secreted isoform. The increase in FGFR-1 mRNA levels in the 22.5-kDa expressing cells was due to enhanced stability of the transcript. Confocal microscopy detected the presence of FGFR-1 at the cell surface whereas secretory isoforms of the receptor were not observed. Reverse transcriptase-polymerase chain reaction did not reveal significant differences in the expression of FGFR-1 variants. In the 22.5-kDa expressing cells exogenous FGF-2 evoked a stronger translocation of the calcium-phospholipid-dependent PEC. These results indicate that the transfected FGF-2 isoforms up-regulated FGFR-1 mRNA and protein. The 22.5-kDa form acted by increasing FGFR-1 mRNA stability enhancing cell responses to exogenous FGF-2.",
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AU - Miquel, Karine

AU - Gaubert, François

AU - Hollande, Etienne

AU - Korc, Murray

AU - Vaysse, Nicole

AU - Clemente, François

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