Increase in globin chains and globin mRNA in erythroleukemia cells in response to hemin

Dabney, Betty J.; Beaudet, Arthur L.

doi:10.1016/0003-9861(77)90092-3

Cited by 70 publications

(14 citation statements)

References 16 publications

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“…5, middle). Consistent with previous reports (10), little or no detectable globin mRNA was present in uninduced MEL cells, but high levels were observed in response to both hemin and DMSO treatment. The same blot was hybridized to a rat tubulin cDNA as a loading control (Fig.…”

supporting

confidence: 92%

Erythroid expression of the heme-regulated eIF-2 alpha kinase.

et al. 1994

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The role of heme-regulated eIF-2a kinase (HRI) in the regulation of protein synthesis in rabbit reticulocytes is well documented. Inhibitors of protein synthesis with properties similar to those of HRI have been described in some nonerythroid cell types, but it has not yet been determined whether these eIF-2o kinase activities are mediated by HRI or one or more as yet uncharacterized kinases. We have studied the expression of mRNA, polypeptide, and kinase activities of HRI in various tissues from both nonanemic and anemic rabbits. Our results indicate that HRI is expressed in an erythroid cell-specific manner. HRI is present in the bone marrow and peripheral blood of both nonanemic and anemic rabbits but not in any of the other tissues tested. HRI mRNA is present at low levels in uninduced mouse erythroleukemic (MEL) cells and human K562 cells and accumulates to higher levels upon induction. The accumulation of HRI mRNA in differentiating MEL cells is dependent upon the presence of heme. The addition of 3-amino-1,2,4-triazole (AT), an inhibitor of heme biosynthesis, to the induction medium markedly reduced HRI mRNA accumulation. Simultaneous addition of hemin and AT to the dimethyl sulfoxide induction medium largely prevented the inhibition of HRI mRNA induction by AT. These findings indicate that HRI is expressed in an erythroid cell-specific manner and that the major physiologic role of HRI is in adjusting the synthesis of globins to the availability of heme.Protein synthesis in intact reticulocytes and their lysates is dependent on the availability of heme. In heme deficiency, inhibition of protein synthesis occurs as a result of the activation of the heme-regulated inhibitor HRI (5,21,23,31). HRI is a cyclic AMP-independent protein kinase which specifically phosphorylates the 38-kDa ot subunit (eIF-2a) of eukaryotic initiation factor 2 (eIF-2) (18,26,28,46). Phosphorylation of eIF-2ao in reticulocyte lysates results in the binding and sequestration of eIF-2B in an eIF2(cP)-eIF-2B complex. The unavailability of eIF-2B, which is required for the exchange of GTP for GDP in the recycling of eIF-2 and the formation of the ternary complex (eIF-2/GTP/Met-tRNAf), results in the cessation of the initiation of protein synthesis (reviewed in references 21, 23, and 31).HRI is present in an inactive form in hemin-supplemented reticulocyte lysates (33, 34) and becomes active under conditions of heme deficiency (33,34) Ehrlich ascites cells which they believe to be distinct from HRI and the double-stranded RNA-dependent eIF-2ot kinase (PKR). Furthermore, increased eIF-2ot phosphorylation has been observed in HeLa and Ehrlich ascites cells under conditions of heat shock and nutrient starvation (11,13,14,39,50), in GH3 pituitary cells treated with calcium-mobilizing agents (45), and in vasopressin-treated rat liver (24). It has not yet been determined whether the eIF-2ot kinase activities measured in these cells are mediated by HRI or one or more as yet uncharacterized eIF-2a kinases.Previous studies using a monoclonal ant...

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confidence: 92%

Erythroid expression of the heme-regulated eIF-2 alpha kinase.

et al. 1994

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“…Furthermore, hemin (ferric chloride protoporphyrin IX) has interesting biological properties such as stimulation ofneurite outgrowth (13), promotion ofadipocyte differentiation (14), and stimulation of globin mRNA accumulation in erythroleukemic cells (15,16). Previously, we demonstrated that hemin increased the levels of functional mRNA for heme oxygenase in cultured pig alveolar macrophages (17) and in rat liver (18), suggesting that hemin acts at the transcriptional level to increase the amount of heme oxygenase.…”

mentioning

confidence: 99%

Cloning and expression of cDNA for rat heme oxygenase.

Shibahara¹,

Müller²,

Taguchi³

et al. 1985

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

412

235

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Two cDNA clones for rat heme oxygenase have been isolated from a rat spleen cDNA library in kgtll by immunological screening using a specific polyclonal antibody. One of these clones has an insert of 1530 nucleotides that contains the entire protein-coding region. To confrm that the isolated cDNA encodes heme oxygenase, we transfected monkey kidney cells (COS-7) with the cDNA carried in a simian virus 40 vector. The heme oxygenase was highly expressed in endoplasmic reticulum of transfected cells. The nucleotide sequence of the cloned cDNA was determined and the primary structure of heme oxygenase was deduced. Heme oxygenase is composed of 289 amino acids and has one hydrophobic segment at its carboxyl terminus, which is probably important for the insertion of heme oxygenase into endoplasmic reticulum. The cloned cDNA was used to analyze the induction of heme oxygenase in rat liver by treatment with CoCl2 or with hemin. RNA blot analysis showed that both CoCl2 and hemin increased the amount of hybridizable mRNA, suggesting that these substances may act at the transcriptional level to increase the amount of heme oxygenase.The microsomal heme oxygenase plays an essential role in physiological heme catabolism (1, 2). Heme oxygenase catalyzes the oxidative degradation of heme to biliverdin (1), which is subsequently converted to bilirubin by biliverdin reductase (3). In the rat, the activity of heme oxygenase is highest in the spleen, where senescent erythrocytes are sequestrated and destroyed (2). Other tissues such as bone marrow and liver also perform this function, especially in hemolytic states and after splenectomy (2, 4). Heme oxygenase is highly inducible by its substrate heme in kidney (2,5), liver (2, 5), and macrophages (6-8). Heme oxygenase is also induced by various other substances such as metal ions (9, 10), endotoxin (11), and bromobenzene (12).We are particularly interested in the induction of heme oxygenase by heme, because heme (ferrous protoporphyrin IX) is an essential component of hemoglobin and of other hemoproteins. Furthermore, hemin (ferric chloride protoporphyrin IX) has interesting biological properties such as stimulation ofneurite outgrowth (13), promotion ofadipocyte differentiation (14), and stimulation of globin mRNA accumulation in erythroleukemic cells (15,16). Previously, we demonstrated that hemin increased the levels of functional mRNA for heme oxygenase in cultured pig alveolar macrophages (17) and in rat liver (18), suggesting that hemin acts at the transcriptional level to increase the amount of heme oxygenase. To understand the molecular mechanisms of induction of heme oxygenase, it is essential to know the structure of the gene for heme oxygenase.In this study, we have isolated cDNA clones for rat heme oxygenase by antibody screening, and we have confirmed that our cDNA actually encodes heme oxygenase by expressing cDNA in monkey kidney cells. We determined the nucleotide sequence of the cloned cDNA and deduced the amino acid sequence of heme oxygenase. MATERIALS AN...

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“…Effects of Hemin and AmLev. Growth of cells from clone 745, the parent of FTG3CI, in hemin-containing medium results in the induction of both globin mRNA and hemoglobin accumulation (14,22). When Me2SO and hemin are applied simultaneously, there is a synergistic effect on hemoglobin accumulation but the level of globin mRNA is no higher than that induced by Me2SO alone.…”

Section: Resultsmentioning

confidence: 92%

“…Other investigators have found that heme can induce increased accumulation of globin mRNA in erythroleukemic cells (14,22). Because Me2SO induces both increased heme biosynthesis and globin mRNA accumulation in these cells, the ability of heme to induce globin mRNA could suggest that it acts as an intermediate in Me2SO-induced globin mRNA accumulation.…”

Section: Resultsmentioning

confidence: 99%

Negative control of hemoglobin production in somatic cell hybrids due to heme deficiency.

Benoff¹,

Bruce²,

Skoultchi³

1978

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

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In somatic cell hybrids formed by the fusion of mouse erythroleukemic cells with mouse primary bone marrow cells, retention of the X chromosome contributed by the bone marrow parent is correlated with inhibition of hemoglobin accumulation in response to dimethyl sulfoxide. The inhibition of hemoglobin accumulation is not due to the absence of globin mRNA. Dimethyl sulfoxide-treated hybrid cells accumulate polyribosomal globin mRNA to levels comparable to those of the parental erythroleukemic cells under the same conditions. Heme, or its precursor delta-aminolevulinc acid, can overcome the effects of the bone marrow X chromosome and induce hemoglobin accumulation in the dimethyl sulfoxide-treated hybrid cells. The data suggest that the X chromosome contributed by the bone marrow cells inhibits hemoglobin production by inhibiting inducible heme biosynthesis, most probably at the step catalyzed by delta-aminolevulinic acid synthetase (EC 2.3.1.37).

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Increase in globin chains and globin mRNA in erythroleukemia cells in response to hemin

Cited by 70 publications

References 16 publications

Erythroid expression of the heme-regulated eIF-2 alpha kinase.

Erythroid expression of the heme-regulated eIF-2 alpha kinase.

Cloning and expression of cDNA for rat heme oxygenase.

Negative control of hemoglobin production in somatic cell hybrids due to heme deficiency.

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