Stimulation of the release of two glycoproteins from mouse 3T3 cells by growth factors and by agents that increase intralysosomal pH

Nilsen-Hamilton, Marit; Hamilton, Richard T.; Allen, W. Ross; Massoglia, S.

doi:10.1016/0006-291x(81)91275-4

Cited by 57 publications

(24 citation statements)

References 17 publications

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“…Nilsen-Hamilton et al (18) have recently shown that preparations of bovine brain fibroblast growth factor (9) stimulate BALB/c-3T3 cells to secrete MEP within 16 h of growth factor addition; DNA synthesis occurs at 12 h. The present study shows that PDGF stimulates intracellular pII (MEP) synthesis within 40 min, long before the onset of DNA synthesis. Pure PDGF stimulated pll (MEP) synthesis at nanogramsper-milliliter concentrations, whereas the growth factors present in plasma did not.…”

Section: Resultsmentioning

confidence: 48%

Identification of a BALB/c-3T3 cell protein modulated by platelet-derived growth factor.

Scher¹,

Dick²,

Whipple³

et al. 1983

Mol. Cell. Biol.

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The platelet-derived growth factor (PDGF) stimulates density-arrested BALB/c-3T3 cells to synthesize a protein (pII; Mr, 35,000) that is constitutively synthesized by spontaneously transformed BALB/c-3T3 (ST2-3T3) cells which do not require PDGF for growth. Antisera against a major excreted protein family (MEP) of retrovirus-transformed cells quantitatively precipitated cellular pII. PDGF-stimulated pII has the same molecular weight, a similar charge, and similar antigenic determinants as authentic MEP isolated from ST2-3T3 or retrovirustransformed cells. MEP represented about 2% of the nonnuclear proteins synthesized by ST2-3T3 cells and 0.3 to 0.6% of the proteins synthesized by PDGFtreated BALB/c-3T3 cells, a three-to sixfold increase over the background. In BALB/c-3T3 cells, less PDGF was required for pII (MEP) synthesis than for DNA synthesis. PDGF induced a selective increase in pII (MEP) within 40 min. Such preferential synthesis was inhibited by brief treatment with actinomycin D, suggesting a requirement for newly formed RNA. The constitutive synthesis of pll (MEP) by ST2-3T3 cells was not inhibited by actinomycin D. Five spontaneously or chemical carcinogen-transformed tumorigenic BALB/c-3T3 cell lines were studied; they neither required PDGF for growth nor responded to it. These cell lines became arrested at confluence with a G1 DNA content. Each of these independently isolated lines synthesized pII (MEP) constitutively. Thus, the synthesis of pII (MEP) may be required, but is not sufficient, for PDGFmodulated DNA synthesis.

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Section: Resultsmentioning

confidence: 48%

Identification of a BALB/c-3T3 cell protein modulated by platelet-derived growth factor.

Scher¹,

Dick²,

Whipple³

et al. 1983

Mol. Cell. Biol.

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“…In contrast to BALB/c 3T3 and SV40-transformed Swiss 3T3 cells, addition of fibroblast growth factor to Swiss 3T3 cells stimulated them to synthesize and secrete MRP (14). To assess relationships between cytoplasmic MRP mRNA content and the ability or inability to secrete MRP, we compared several permanent mouse cell lines by Northern gel analysis, as described elsewhere (4; D. R. Edwards, C. L. J. Parfett, and D. T. Denhardt, Mol.…”

mentioning

confidence: 85%

“…This conclusion was supported in a study of transcriptional regulation of serum-induced RNAs from Swiss 3T3 fibroblasts in which both the level of MRP mRNA within the nucleus (Edwards et al, in press) and the amount of nuclear runoff transcription increased after serum stimulation of Swiss 3T3 cells (D. R. Edwards, unpublished observation). Furthermore, the induction of MRP mRNA and cellular proliferative responses are separable, since this species was undetectable in the cytoplasm of serum- Although MRP mRNA was inducible in both Swiss 3T3 and BALB/c 3T3 mouse cells, the protein was not detected in BALB/c 3T3 cells after stimulation by calf serum or purified growth factors (12,14), pointing to possible translational or postranslational regulatory mechanisms that operate differently in these cell lines. In this regard it has been shown that lysosomal degradation of newly synthesized MRP plays a major role in determining the quantity of MRP secreted by Swiss 3T3 cells into the culture medium (14; Nilsen-Hamilton et al, submitted for publication).…”

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confidence: 95%

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Characterization of a cDNA clone encoding murine mitogen-regulated protein: regulation of mRNA levels in mortal and immortal cell lines.

Parfett¹,

Hamilton²,

Howell³

et al. 1985

Mol. Cell. Biol.

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Mitogen-regulated protein (MRP) is secreted by certain immortal murine cell lines (Swiss 3T3, BNL) stimulated with serum or particular growth factors. We have identified a cDNA clone that encodes part of the protein and have confirmed that MRP is closely related to, if not identical to, the prolactin-related protein designated proliferin. MRP is not produced by primary mouse embryo fibroblasts to nearly the same extent as it is produced by many immortal or transformed lines. Control of expression of this protein by growth factors is achieved both by regulating the extent of transcription and by regulating the processing of the protein.Mitogen-regulated protein (MRP) was described by Nilsen-Hamilton et al. (12) (Mr, 30,000 to 38,000) released by Swiss mouse 3T3 cells some 18 to 28 h after stimulation with growth factors (serum, fibroblast growth factor, epidermal growth factor); platelet-derived growth factor and the tumor promoter 12-tetra-decanoylphorbol-13-acetate were shown subsequently to be effective at stimulating release also. Evidence for glycosylation includes the facts that treatment with endoglycosidase H reduces the apparent molecular weight substantially and that the protein synthesized in the presence of tunicamycin has a lower Mr of about 22,000. An antiserum raised against purified MRP has been used to identify the protein unambiguously.Not only mitogens, but also substances that increase intralysosomal pH (NH4CI, chloroquine, and the ionophores monensin and nigericin) enhance the production of MRP (14). This suggests that MRP levels are, in part, regulated at the level of protein degradation, since these latter substances inhibit degradation of lysosomal proteins. In agreement with this is the observation that the ionophore-stimulated release of MRP is not blocked by actinomycin D, in contrast to growth factor-induced release (M. Nilsen-Hamilton and R. T. Hamilton, unpublished data). Proteins known to be secreted by fibroblasts include plasminogen activator, collagenase, fibronectin, procollagen, and major excreted protein (13). Proteins for which there is evidence (in other cell types) that the secreted quantities are regulated by intracellular degradation include prolactin, parathormone, and procollagen (2).We have used an antiserum raised against MRP (R. T. Hamilton and M. Nilsen-Hamilton unpublished data) to screen a Xgtll expression library (18) for clones able to code for one or more epitopes found on MRP. Mouse BNL cells (15) were used as the source of poly(A) mRNA for cloning, since among the cell lines tested it produced the most MRP. * Corresponding author.To provide initial enrichment for poly(A) mRNA molecules coding for MRP, the cytoplasmic poly(A) mRNA extracted from a growing population of mouse BNL cells was fractionated by velocity sedimentation in a sucrose gradient containing formamide. We recovered the RNA in separate pools and examined it by in vitro translation and immunoprecipitation to identify the fraction most enriched for MRP mRNA. The cDNA synthesized from that fra...

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“…For-fibroblasts, serum and purified growth factors selectively stimulate the synthesis of proteins that are found intracellularly (5-7). However, it has been found that the most dramatic selective effects of growth factors on fibroblast protein synthesis are in proteins that are secreted (8)(9)(10). It has been proposed that these secreted, mitogen-induced proteins (MIPs) are involved in coordinating cell growth during tissue regeneration or embryogenesis (2).…”

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confidence: 99%

Rapid selective effects by a growth inhibitor and epidermal growth factor on the incorporation of [35S]methionine into proteins secreted by African green monkey (BSC-1) cells.

Nilsen-Hamilton¹,

Holley²

1983

Proc. Natl. Acad. Sci. U.S.A.

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Because growth stimulators selectively induce certain secreted proteins, we asked whether growth inhibitors, which antagonize the effects of growth factors on DNA synthesis, act independently to induce other proteins or whether they counteract the effects of growth factors on protein induction. To answer this question, we have worked with an epithelial cell line (BSC-1) derived from African green monkey kidney cells. BSC-1 cells are stimulated to divide by epidermal growth factor (EGF) and are inhibited in DNA synthesis and growth by an inhibitor that is secreted by confluent cultures of the same cells (11).We have found that BSC-1 cells respond to the purified protein growth inhibitor by selectively increasing the rate at which they synthesize and secrete a protein with an approximate Mr of 48,000. These same cells respond to EGF, a growth stimulus, by selectively increasing the rates at which they synthesize and secrete three other proteins with approximate Mrs of 28,000, 59;000, and 62,000. The effects of the growth inhibitor and EGF are additive both in the induction of secreted proteins and on DNA synthesis.MATERIALS AND METHODS Materials. The high molecular weight growth inhibitor was purified by chromatography through Bio-Gel P-60 followed high-performance liquid chromatography as described (11).[S]Methionine was from Amersham, actinomycin D was from Calbiochem, and aprotinin (Trasylol) was from FBA Pharmaceuticals, New York. Cell Culture. BSC-1 cells were cultured as described (12) 5636The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S.C. §1734 solely to indicate this fact.

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Stimulation of the release of two glycoproteins from mouse 3T3 cells by growth factors and by agents that increase intralysosomal pH

Cited by 57 publications

References 17 publications

Identification of a BALB/c-3T3 cell protein modulated by platelet-derived growth factor.

Identification of a BALB/c-3T3 cell protein modulated by platelet-derived growth factor.

Characterization of a cDNA clone encoding murine mitogen-regulated protein: regulation of mRNA levels in mortal and immortal cell lines.

Rapid selective effects by a growth inhibitor and epidermal growth factor on the incorporation of [35S]methionine into proteins secreted by African green monkey (BSC-1) cells.

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