Stimulation of DNA synthesis, cell multiplication, and ornithine decarboxylase in 3T3 cells by multiplication stimulating activity (MSA)

Nissley, S. Peter; Passamani, John; Short, Patricia A.

doi:10.1002/jcp.1040890305

Cited by 51 publications

(18 citation statements)

References 22 publications

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“…It was identified during the purification by its ability to stimulate the incorporation of [3H]thymidine into the DNA of chick-embryo fibroblasts [8]. Following Dowex 50 chromatography, Sephadex G-75 gel filtration in 1 M acetic acid, and preparative acrylamide gel electrophoresis at pH 2.7 in 9 M urea, fractions were obtained in which MSA appeared to be nearly homogeneous by both charge and size criteria [9,17]. Analytical disc acrylamide electrophoresis at pH 2.7 in 9 M urea and at pH 10 without denaturants gave a single protein band that stimulated [3H]thymidine incorporation into chick embryo fibroblasts [9,17].…”

Section: Polypeptidesmentioning

confidence: 99%

“…Following Dowex 50 chromatography, Sephadex G-75 gel filtration in 1 M acetic acid, and preparative acrylamide gel electrophoresis at pH 2.7 in 9 M urea, fractions were obtained in which MSA appeared to be nearly homogeneous by both charge and size criteria [9,17]. Analytical disc acrylamide electrophoresis at pH 2.7 in 9 M urea and at pH 10 without denaturants gave a single protein band that stimulated [3H]thymidine incorporation into chick embryo fibroblasts [9,17]. When *2SI-labeled MSA (see below) that had been reduced and alkylated in 6 M guanidine HCI was chromatographed on a 6 % agarose column equilibrated with 6 M guanidine HCI, the radioactivity eluted as a single peak corresponding to a molecular weight of 8700 [9].…”

Section: Polypeptidesmentioning

confidence: 99%

“…Chick-embryo fibroblast cultures were established from 12-day-old embryo carcasses and grown in ET medium (Temin's modified Eagle's medium with 20% by volume tryptose phosphate broth [8,17,19] Cultures of human fibroblasts were developed from punch biopsies of forearm skin from normal volunteers. Fibroblasts were grown at 37 "C in 95% air/ 5 : ; C 0 2 in modified Eagle's minimum essential medium supplemented with 10 mM Hepes, pH 7.4, 207; fetal calf serum, and 5 mg/100 ml neomycin.…”

Section: Cell Cultivationmentioning

confidence: 99%

“…After 3 days, MSA, somatomedin A, somatomedin B or media alone were added in E medium. After 12 h, the cultures were pulsed for 1 h with [3H]thymidine, 0.4 pCi/2 ml, and the incorporation of radioactivity into acid-precipitable material determined in duplicate dishes [17,19].…”

Section: [3h]thymidine Incorporation Inio the Dna Of Chick Embryo Fibmentioning

confidence: 99%

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Purified Human Somatomedin A and Rat Multiplication Stimulating Activity

Rechler

Fryklund

Nissley

et al. 1978

European Journal of Biochemistry

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We have compared the mitogenic activity and reactivity in receptor binding assays of somatomedin A and multiplication-stimulating activity (MSA), two acid-soluble polypeptides of molecular weight 7-10000 that possess weak intrinsic insulin-like metabolic activity. Somatomedin A was purified to homogeneity from human plasma, and MSA was purified to homogeneity from the conditioned culture media of a cell line, BRL 3A, derived from rat liver. The two peptides stimulated DNA synthesis in chick embryo fibroblasts 3 -4-fold and have superimposable doseresponse curves. Addition of either peptide to chick embryo fibroblasts plated in serum-free medium increased the cell number by 150 in 4 days, compared with an increase of 25 % in control cultures receiving no additions. The two peptides also stimulated DNA synthesis in human fibroblasts in culture 3 -4-fold.Specific receptors for MSA and/or somatomedin A could be demonstrated in intact cells or membranes from chick embryo fibroblasts, human fibroblasts, human placenta, rat liver, and the BRL 3A2 cell line, a subclone of the line that produces MSA. Unlabeled MSA and somatomedin A inhibited the binding of '251-labeled MSA and '251-labeled somatomedin A to each of these receptors with comparable potency. In chick embryo fibroblasts, human fibroblasts, and human placental membranes, the binding of both radioactive ligands also was inhibited by insulin, consistent with the interpretation that lz5I-1abeled MSA and '251-labeled somatomedin A were binding to the same receptor. By contrast, in the BRL 3A2 cell line, insulin inhibited the binding of '251-labeled somatomedin A, but not the binding of '251-labeled MSA, suggesting that the two labeled peptides were binding to different receptors in this cell line. Moreover, '251-labeled MSA, but not '251-labeled somatomedin A, bound specifically to rat liver plasma membranes. These results indicate that human somatomedin A and rat MSA are closely related, but not identical, peptides. They have similar mitogenic activity for cultured fibroblasts, and cross-react with specific cell receptors with comparable potency. The two radioactively labeled peptides, however, can be distinguished by certain cell surface receptors for somatomedin-like peptides.The growth-promoting actions of growth hormone on its target tissues are thought to be mediated by substances in plasma that have been designated somatomedins [ 11. Several growth hormone-dependent polypeptides with properties of somatomedins have Abhreviarions.

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Section: Polypeptidesmentioning

confidence: 99%

Section: Polypeptidesmentioning

confidence: 99%

Section: Cell Cultivationmentioning

confidence: 99%

Section: [3h]thymidine Incorporation Inio the Dna Of Chick Embryo Fibmentioning

confidence: 99%

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Purified Human Somatomedin A and Rat Multiplication Stimulating Activity

Rechler

Fryklund

Nissley

et al. 1978

European Journal of Biochemistry

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“…Independently, an activity was identified that lowered glucose levels even in the presence of anti-insulin antibodies (nonsuppressible insulin like activity, NSILA) (Oelz et al 1970;Megyesi et al 1974;Rechler et al 1974). Additionally, a protein synthesized by the liver, mitogenic for fibroblasts, and termed multiplication stimulating activity (MSA) was identified (Nissley et al 1976). With their purification and cloning, all of these activities were found to be attributable to IGF-I (Jansen et al 1983;Ullrich et al 1984) and IGF-II (Bell et al 1984;Ullrich et al 1984;Whitfield et al 1984).…”

mentioning

confidence: 99%

IGF-I is required for normal embryonic growth in mice.

Powell-Braxton¹,

Hollingshead²,

Warburton³

et al. 1993

Genes & Development

718

465

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IGF-I is a pleiotropic hormone reported to affect linear growth, glucose metabolism, organ homeostasis, and the immune and neurologic systems. In contrast to IGF-II, IGF-I is expressed at low levels embryonically and has been thought to be more important for postnatal growth and development. To investigate the role of IGF-I in normal development we generated mice with an inactive IGF-I gene by homologous recombination in ES cells. Heterozygous mice are healthy and fertile, but they are 10-20% smaller than wild-type littermates and have lower than normal levels of IGF-I. The size reduction is attributable to a decrease in organs and muscle and bone mass. However, all tissues appear histologically normal. At birth homozygous mutant mice (IGF-I -/-) are <60% body weight of wild type. Greater than 95% of IGF-I -/-pups die perinatally. Histopathology is characterized by underdevelopment of muscle tissue. Lungs of late embryonic and neonates also appeared less organized with ill-defined alveolae. IGF-I appears to be essential for correct embryonic development in mice.

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Physiological aspects of ornithine decarboxylase

Bachrach

1984

Cell Biochemistry & Function

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Physiological aspects of ornithine decarboxylaseOrnithine decarboxylase (ODC, L-ornithine carboxylyase, EC 4.1.1.17) is the rate limiting enzyme in the biosynthesis of polyamines. Its occurrence in animal tissue was reported independently by three laboratories in 196813, and its activity increases in various growing tissues and organs. During recent years, this enzyme, and its regulation, have attracted considerable attention and available information has been summarized in several reviewsk7. It is now well established that virtually all hormones cause the increase of ODC activity in appropriate target tissues (Tubte 1). In many cases, this increase is mediated by cyclic AMP ( Table 2). Properties of ornithine decarboxylaseThe properties of ornithine decarboxylase are dealt with in the previous Paper by A. M. Kaye (p 2). Regulation of ornithine decarboxylase activityOne of the typical features of ODC is its short half-life, which ranges from 15 to 45 min. This is the shortest half-life of any enzyme in eukaryotes. To account for such rapid changes in activity, several mechanisms have been proposed6'. The three major mechanisms are depicted in Table 3. AntizymeIn 1976, Canellakis and coworkers6' described an M, 26 000 protein, which was formed upon treating cells in culture with 5-10 mM putrescine. This protein, which was designated as antizyme, formed complexes with ODC and neutralized its activity. The complex could be dissociated by ammonium sulphate or by relatively high salt concentration of 250 m~~~. The antizyme was first detected in H-35 rat hepatoma cells, L-1210 mouse leukaemia cells, N-18 mouse neuroblastoma cells, rat liver, and subsequently, also in many other cell linesm and also in bacteria65. It is noteworthy that the antizyme appeared to be ubiquitous and a bacterial antizyme could neutralize a mammalian a n t i~y r n e~~.One difficulty in studies on antizymes has been the lack of an appropriate method for the assay of the intracellular amount of enzyme-antizyme complexes. Fujita et al. have recently described the presence of an 'antizyme inhibitor' in rat liver66. This protein, which resembles ODC in its molecular weight, could replace antizyme in a complex with ODC, and reactivate the latter. It is tempting to speculate that this antizyme inhibitor is a modified form of ODC (such as a phosphorylated or transaminated form). This assumption should be confirmed by peptide and immunological analyses. The 'antizyme inhibitor' could eventually be used for the assay of the amount of inactive ODC in a complex with antizyme". It is not certain why the polyamine-dependent protein kinase of P. polycephalum required both calcium ions and calmodulin for phosphorylation, while that of the hepatoma cells only required calmodulin. This discrepancy may be explained by the different source of the enzymes and by the degree of purity. One enzyme was of fungal origin, while the other was a mammalian one. Moreover, the P. polycephalum enzyme was derived from nuclear extract, and autophosphorylation was measured. The ...

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Stimulation of DNA synthesis, cell multiplication, and ornithine decarboxylase in 3T3 cells by multiplication stimulating activity (MSA)

Cited by 51 publications

References 22 publications

Purified Human Somatomedin A and Rat Multiplication Stimulating Activity

Purified Human Somatomedin A and Rat Multiplication Stimulating Activity

IGF-I is required for normal embryonic growth in mice.

Physiological aspects of ornithine decarboxylase

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