Erythroid cell differentiation: murine erythroleukemia cell variant with unique pattern of induction by polar compounds.

Ohta, Yoshikuni; Tanaka, Masao; Terada, Masaaki; Miller, Orlando J.; Bank, Arthur; Marks, Paul A.; Rifkind, Richard A.

doi:10.1073/pnas.73.4.1232

Cited by 97 publications

(47 citation statements)

References 12 publications

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“…Strain 745-A, which is infected with Friend virus complex, was kindly provided by Charlotte Friend and has been maintained in culture in our laboratory as described elsewhere (14). Isolation of the Me2SO-sensitive cell line (DS19) and resistant cell line (DR10) was described by Ohta et al (15). Cell concentration and proportion of hemoglobin-containing (benzidine-reactive) cells were determined as previously described (14).…”

mentioning

confidence: 99%

Transient inhibition of initiation of S-phase associated with dimethyl sulfoxide induction of murine erythroleukemia cells to erythroid differentiation.

Terada¹,

Fried²,

Nudel³

et al. 1977

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

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108

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The murine erythroleukemia cell (MELC) line in suspension culture can be induced to differentiate to erythroid cells by various compounds, including dimethyl sulfoxide (Me2SO). Analysis of the cell cycle, during differentiation induced by Me2SO, using thymidine incorporation, thymidine labeling index, and relative DNA content per cell as measured by flow microfluorometry, demonstrates a transient inhibition of entry of (2), accumulation of globin mRNAs (3-6), a and f globin synthesis (7), increase in heme synthesis (8), synthesis of erythrocyte-specific proteins (9), loss in capacity for cell division (2, 10), and the appearance of erythrocytespecific membrane proteins (11).The rate of DNA synthesis, proportion of cells in S-phase, and the pattern of DNA accumulation have been measured and compared in MELC grown in the presence and absence of Me2SO and other inducing agents, e.g., butyric acid (12) and dimethylacetamide (13). DNA synthesis has been measured using incorporation of isotopically labeled thymidine in pulse-labeling experiments; cells in S-phase were assayed by autoradiography of thymidine pulse-labeled cells; DNA accumulation per cell was determined using propidium iodide staining with flow microfluorometric analysis. By MATERIALS AND METHODS Cells. Strain 745-A, which is infected with Friend virus complex, was kindly provided by Charlotte Friend and has been maintained in culture in our laboratory as described elsewhere (14). Isolation of the Me2SO-sensitive cell line (DS19) and resistant cell line (DR10) was described by Ohta et al. (15). Cell concentration and proportion of hemoglobin-containing (benzidine-reactive) cells were determined as previously described (14).Pulse Labeling with Thymidine. DNA synthesis was determined as incorporation of [3Hjthymidine (20 Ci/mmol, New England NuclearCorp.) into cells (2 X 105 cells/per ml) incubated for 20 min at 370 at a concentration of 20 ,Ci/ml of culture medium. Thymidine pulse labeling in this manner was performed on aliquots removed from culture at the times indicated for each experiment. Following incubation with radioisotope, cells were washed with phosphate-buffered saline, pH 7.4, containing 2 mM thymidine, and precipitated with 10% trichloroacetic acid. Precipitates were collected on Millipore filters and their radioactivity was measured in Aquasol. A separate aliquot of cell suspension was removed from the culture and, washed with the same phosphate-buffered saline, and smears were prepared employing the cytocentrifuge. These slides were processed for radioautography (16

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confidence: 99%

Transient inhibition of initiation of S-phase associated with dimethyl sulfoxide induction of murine erythroleukemia cells to erythroid differentiation.

Terada¹,

Fried²,

Nudel³

et al. 1977

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

Self Cite

108

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“…A Sequenase® 2.0 kit was obtained from U. S. Biochemical Corp. C-18 reverse-phase LUNA column was purchased from Phenomenex. N23 murine erythroleukemia is a subclone of DS19 cells (19), a murine erythroleukemia cell line obtained as described previously (20).…”

Section: Methodsmentioning

confidence: 99%

“…Assay of Calpain Activator Activity-Calpain activator activity was measured by adding the appropriate amounts of the activator sources to the routine calpain assay mixture, which contained 50 mM sodium borate buffer, pH 7.5, 5 M CaCl 2 , 2 mg/ml of denatured globin, and 0.01 nmol of erythrocyte calpain (19). 1 unit of calpain activator activity is defined as the amount that elicits 1 unit of calpain activity in the presence of 5 M Ca 2ϩ (7).…”

Section: Methodsmentioning

confidence: 99%

“…The primers for PCR were selected externally to the translated sequence of mouse brain cDNA for ACBP (accession number X61431). The sense primer was 5Ј-CTTGATTGCTCCTGCTTCTG-3Ј (nucleotides [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20]; the antisense primer was 5Ј-TATGTCCTCACAGGTCACACG-3Ј (complementary to nucleotides 353-373). PCR was carried out by using Amplitaq DNA polymerase and by performing 30 cycles of amplification as described previously (24).…”

Section: Methodsmentioning

confidence: 99%

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Acyl-CoA-binding Protein Is a Potent m-Calpain Activator

Melloni

Averna

Salamino

et al. 2000

Journal of Biological Chemistry

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Acyl-CoA-binding protein, a 20-kDa homodimer that exerts many physiological functions, promotes activation of the classic calpain forms, most markedly that of the m-isozyme. This protein factor was purified from rat skeletal muscle and was also expressed in Escherichia coli. Both native and recombinant acyl-CoA-binding proteins show the same molecular properties and an identical capacity to decrease the [Ca 2؉ ] required for m-calpain activity. The binding of long-chain acyl-CoAs to acyl-CoA-binding protein does not modify the activating effect on calpains. Acyl-CoA-binding protein seems to be involved in the m-calpain regulation process, whereas the previously identified UK114 activator is a specific modulator of -calpain. Acyl-CoA-binding protein is proposed as a new component of the Ca 2؉ -dependent proteolytic system. A comparative analysis among levels of classic calpains and their activator proteins is also reported.How the calcium-dependent proteolytic system is activated is an intriguing question in terms of understanding its general properties, physiological functions, and involvement in the occurrence of tissue damage (1-4). Calpains, the enzymatic components of the system, are primarily regulated by calcium ions. The binding of Ca 2ϩ to calmodulin-like domains of the proteinases induces a conformational change that constitutes the initial limiting step of the overall sequential activation process (5). In this new conformation calpains undergo limited autoproteolysis that irreversibly removes the molecular constraints responsible for stabilizing the native inactive form (1-4). In their autoproteolyzed active forms, calpains acquire the highest affinity for calpastatin, which becomes the only negative modulator of calpain activity (6). In rat brain we have recently identified a protein factor, called UK114, which specifically interacts with -calpains and accelerates their activation steps, increasing the affinity for calcium ions (7). The reduced time of calpain activation is consistent with the proposed involvement of the Ca 2ϩ -dependent proteolytic system in specific signal transduction pathways, resulting, for instance, in granule secretion in various cell types (8) or in the conversion of protein kinase C into an autoproteolyzed form produced during the differentiation of murine erythroleukemia cells (9).The rat brain -calpain activator shows a strict specificity for all -calpain isoforms, whereas it has no effect on the classic m-calpains (10), the activation of which requires a calcium concentration at least two orders of magnitude higher than that required by -calpain. Despite reports indicating that free calcium ions can be accumulated in specific cell regions at high micromolar concentrations (11), the calcium requirement for m-calpain activation can apparently be satisfied only during tissue necrosis, a situation far removed from physiological conditions (12). We now report that a highly conserved protein, called acylCoA-binding protein (ACBP) 1 (13-15), shows a potent m-calpain-activating pr...

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“…Nonequilibrium pH gradient gel electrophoresis and sodium dodecyl sulfatepolyacrylamide gel electrophoresis of total in vivo protein was performed as described previously (5 (20,26), an increase in the absolute rate of globin gene transcription (8), and a decline in cellular protein content (26). Although the rate of total protein synthesis undergoes a general reduction in induced cells, the synthesis of some proteins remains constant, whereas the synthesis of others increases (20). The virtual de novo synthesis of globin (8,9) and the dramatic increase in the mRNAs for enzymes of the heme biosynthetic pathway (10) in the midst of declining total protein synthesis are excellent examples of specific gene control.…”

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confidence: 99%

Expression of a gene for mouse eucaryotic elongation factor Tu during murine erythroleukemic cell differentiation.

Roth¹,

Bragg²,

Corrias³

et al. 1987

Mol. Cell. Biol.

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The eucaryotic elongation factor Tu (eEF-Tu) is a single polypeptide with an approximate M, of 53,000.During protein synthesis eEF-Tu promotes the binding of aminoacyl-tRNA to the ribosome. To study the expression of the gene(s) for this factor, a genomic clone was isolated that contains a mouse eEF-Tu gene. We screened a phage genomic library with a synthetic oligonucleotide probe complementary to a region of the Saccharomyces cerevisiae and Artemia sp. eEF-Tu genes which codes for an area that is highly conserved between both yeast and Artemui sp. eEF-Tu. From approximately 75,000 phage plaques we obtained five isolates with apparently identical inserts. All five clones contained a 3.8-kilobase EcoRI fragment that hybridized to additional oligonucleotide probes corresponding to different conserved regions of eEF-Tu. We sequenced the 5' end of one genomic clone and determined the length of the cloned fragment that was protected by eEF-Tu mRNA in Si nuclease protection assays. A quantitative SI nuclease protection assay was used to compare the relative steady-state levels of eEF-Tu mRNA in total RNA isolated from hexamethylenebisacetamide-induced muriniP erythroleukemia cells. The results show a dramatic reduction in the steady-state level of eEF-Tu mRNA as differentiation proceeds. A similar reduction in transcription of eEF-Tu mRNA was observed in isolated nuciei. Finally, we examined the in vivo synthesis of eEF-Tu during differentiation and found that it declined in a manner parallel to the decline in the steady-state level of eEF-Tu mRNA. In addition, we have isolated and sequenced a cDNA clone for mouse eEF-Tu. The derived amino acid sequence is compared with sequences from other eucaryotes.

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Erythroid cell differentiation: murine erythroleukemia cell variant with unique pattern of induction by polar compounds.

Cited by 97 publications

References 12 publications

Transient inhibition of initiation of S-phase associated with dimethyl sulfoxide induction of murine erythroleukemia cells to erythroid differentiation.

Transient inhibition of initiation of S-phase associated with dimethyl sulfoxide induction of murine erythroleukemia cells to erythroid differentiation.

Acyl-CoA-binding Protein Is a Potent m-Calpain Activator

Expression of a gene for mouse eucaryotic elongation factor Tu during murine erythroleukemic cell differentiation.

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