Human α-globin gene expression following chromosomal dependent gene transfer into mouse erythroleukemia cells

Deisseroth, Albert; Hendrick, Don

doi:10.1016/0092-8674(78)90082-x

Cited by 102 publications

(72 citation statements)

References 13 publications

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“…Chromosomal analysis and APRT determinations were done as described (11,12). For the analysis of the chromosomal proteins, a suspension of 150 ml of these cells was incubated at a concentration of2-5 X 105 cells per ml with 25 ,uCi (1 Ci = 3.7 Abbreviation: APRT, adenine phosphoribosyltransferase (AMP: pyrophosphate phosphoribosyltransferase, EC 2.4.2.7).…”

Section: Methodsmentioning

confidence: 99%

“…The hybrid cell line 323-4 was isolated in F 12 medium containing 0.002 M alanosine, 10% (vol/vol) fetal calfserum, 1 ,uM ouabain, and adenine as the only purine source (8-11). This selective system is lethal for the human parent and promotes the growth of hybrid cells that have retained human chromosome 16, which contains the human a-globin gene (9) and the adenine phosphoribosyltransferase (APRT; AMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.7) gene (APRT) as reported (10).Chromosomal analysis and APRT determinations were done as described (11,12). For the analysis of the chromosomal proteins, a suspension of 150 ml of these cells was incubated at a concentration of2-5 X 105 cells per ml with 25 ,uCi (1 Ci = 3.7 Abbreviation: APRT, adenine phosphoribosyltransferase (AMP: pyrophosphate phosphoribosyltransferase, EC 2.4.2.7).…”

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Expression of human and mouse nonhistone chromosomal proteins in hybrid mouse erythroleukemia cells containing a single human chromosome.

Bode¹,

Deisseroth²,

Hendrick³

1981

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

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were distinct in the mouse and human parents and that hybrids which retain primarily mouse chromosomes express the mouse forms of H2B, whereas hybrids which retain primarily human chromosomes contain the human but not the mouse-specific H2B. Lyderson et al. (6) studied the nonhistone chromosomal proteins of a series of human-Chinese hamster hybrids which had retained a majority of Chinese hamster chromosomes (6). Only 10% ofthe human nonhistone chromosomal proteins were unique from those of the Chinese hamster parent in the twodimensional electrophoresis system used, and only one of the seven hybrid cells studied contained an electrophoretic species that comigrated with any of the nonhistone chromosomal proteins of the human parent.Using two-dimensional gel electrophoresis (7), we have identified 400 and 280 nonhistone chromosomal proteins in the mouse and in the human parental cell lines, respectively. Of the 280 nonhistone chromosomal proteins found in the human cells, 70 were not present in the mouse erythroleukemia cells studied. If the genes for these human nonhistone chromosomal proteins were distributed equally among 46 human chromosomes, one would anticipate that the genes for one to two such human specific nonhistone chromosomal proteins would be associated with each human chromosome. In fact, five of the six hybrid cell lines that contained human chromosome 16 did indeed contain a single Mr 65,000 nonhistone chromosomal protein ofpI 6.2, which was not found in the parent mouse cell but which comigrated with a similar protein found in the human parent cell line used for fusion and in two other human cell lines studied. These studies showed, therefore, that nonhistone chromosomal proteins that are absent in mouse cells and that comigrate with human nonhistone chromosomal proteins can be found in mouse-human hybrid cells containing a majority of mouse chromosomes. MATERIAL AND METHODSCells. Fusion of 1 X 108 2292 cells and 1 x 107 179 MEL cells was performed in the presence of 40% (vol/vol) polyethylene glycol and 10% (vol/vol) dimethyl sulfoxide as described (8). The hybrid cell line 323-4 was isolated in F 12 medium containing 0.002 M alanosine, 10% (vol/vol) fetal calfserum, 1 ,uM ouabain, and adenine as the only purine source (8-11). This selective system is lethal for the human parent and promotes the growth of hybrid cells that have retained human chromosome 16, which contains the human a-globin gene (9) and the adenine phosphoribosyltransferase (APRT; AMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.7) gene (APRT) as reported (10).Chromosomal analysis and APRT determinations were done as described (11,12). For the analysis of the chromosomal proteins, a suspension of 150 ml of these cells was incubated at a concentration of2-5 X 105 cells per ml with 25 ,uCi (1 Ci = 3.7 Abbreviation: APRT, adenine phosphoribosyltransferase (AMP: pyrophosphate phosphoribosyltransferase, EC 2.4.2.7).

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Expression of human and mouse nonhistone chromosomal proteins in hybrid mouse erythroleukemia cells containing a single human chromosome.

Bode¹,

Deisseroth²,

Hendrick³

1981

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

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“…Introduction of part of human chromosome 11 or all of 16 into MELCs leads to induced expression of the ~-or a-globin genes at levels comparable with those of the endogenous mouse genes (Deisseroth and Hendrich 1979;Nandi et al 1988).…”

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confidence: 99%

Induction by HMBA and DMSO of genes introduced into mouse erythroleukemia and other cell lines by transient transfection.

Campbell

Kulozik²,

Woodham³

1990

Genes Dev.

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We have found rapid induction of various genes, including human globin genes, in response to hexamethylene bisacetamide (HMBA) and dimethyl sulfoxide (DMSO) in transiently transfected cells. In mouse erythroleukemia cells (MELCs), this effect is detected within 1 hr of exposure of the cells to inducer before the endogenous mouse globin genes are induced. It does not require protein synthesis and is reversed if the inducer is removed. This and other evidence suggest that the mechanism involves a change in activity of a factor intimately involved with transcription, probably as a result of post-translational modification. As such, it may represent an early triggering event in terminal differentiation, and its relevance to the expression of human globin genes in stable transfectants and to induction of the mouse globin genes is discussed. Other cell lines (K562 and NSO) also show this response, which may therefore involve a ubiquitous mechanism. We also found that HMBA depresses the expression of endogenous globin genes in K562, the opposite of this differentiation inducer's effect on MELC.

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“…MEL cells (aprt mutant [10]) were grown in Dulbecco's modified Eagle medium (Irvine Scientific) supplemented with 10% fetal calf serum (HyClone Laboratories, Inc.) and penicillin G (100 U/ml)-streptomycin sulfate (100 ,ug/ml). Cells were transfected by the DEAE-dextran method and, where indicated, induced to differentiate exactly as described previously (7).…”

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Discrimination among potential activators of the beta-globin CACCC element by correlation of binding and transcriptional properties.

Hartzog¹,

Myers²

1993

Mol. Cell. Biol.

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Adult K-globin-like promoters contain a cis-acting element, CCACACCC, that is conserved across species and is required for wild-type levels of transcription. We have studied the contribution of this element and proteins that interact with it to activate 13-globin transcription. We found that an erythroid-like cell line, MEL, contains several proteins that specifically bind the CACCC element. By comparing the DNA-binding properties of promoters with mutations in the CACCC element with the transcriptional activities of these mutant promoters, we found that two CACCC-binding proteins did not bind to mutant promoters that direct decreased levels of transcription. One of these proteins is the transcriptional activator Spl, and the other we have designated CACD (CACCC-binding species D). We subjected CACD to a binding site selection procedure and obtained high-affinity CACD binding sites that are identical to that of the 13-globin CACCC element. This result, combined with our finding that CACD binds the CACCC element with a higher affinity than does Spl, argues that the CACCC element is a target of CACD rather than Spl. The strategy of correlating the results of a binding site selection experiment with those of in vivo expression and in vitro binding studies may allow evaluation of the relative potential of different proteins to activate transcription through a single cis-acting site.The adult P-globin-like promoters contain several ele-

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Human α-globin gene expression following chromosomal dependent gene transfer into mouse erythroleukemia cells

Cited by 102 publications

References 13 publications

Expression of human and mouse nonhistone chromosomal proteins in hybrid mouse erythroleukemia cells containing a single human chromosome.

Expression of human and mouse nonhistone chromosomal proteins in hybrid mouse erythroleukemia cells containing a single human chromosome.

Induction by HMBA and DMSO of genes introduced into mouse erythroleukemia and other cell lines by transient transfection.

Discrimination among potential activators of the beta-globin CACCC element by correlation of binding and transcriptional properties.

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