Expression of Transfected Genes

Chao, Moses V.

doi:10.1007/978-1-4684-5167-2_8

Gene Transfer 1986

DOI: 10.1007/978-1-4684-5167-2_8

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Expression of Transfected Genes

Moses V. Chao

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1987

1990

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Cited by 4 publications

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“…Although the induction of 7-globin RNA expression after transfer to MEL cells has been well documented (Anagnou et al, 1986;Charnay & Henry, 1986;Chao, 1986;Stoeckert et al, 1987), other studies that used a variant of MEL cells deficient in thymidine kinase observed induction of /3-globin but not 7-globin RNA in a majority of the clones studied •This work was supported in part by NIH Grants DK-31232, and by a fellowship to M.W.R. from the Cooley's Anemia Foundation.…”

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confidence: 95%

Expression of the human .gamma.-globin gene after retroviral transfer to transformed erythroid cells

Rixon¹,

Harris²,

Gelinas³

1990

Biochemistry

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Regulation of the human fetal (gamma) globin gene and a series of mutant gamma-globin genes was studied after retroviral transfer into erythroid cells with fetal or adult patterns of endogenous globin gene expression. Steady-state RNA from a virally transferred A gamma-globin gene with a normal promoter increased after induction of erythroid maturation of murine erythroleukemia cells and comprised from 2% to 23% of the mouse beta maj-globin RNA level. RNA expression from the virally transferred A gamma-globin gene comprised 23% of the endogenous G gamma- + A gamma-globin expression in K 562 cells after treatment with hemin. Expression from a virally transferred gamma- or beta-globin gene exceeded endogenous gamma- or beta-globin expression by a factor of 6 or more in the human erythroleukemia line KMOE, in which the endogenous globin genes are weakly inducible. In these experiments, no difference in expression was observed between the gene with the normal promoter and an A gamma-globin gene with a point mutation in its promoter (-196 C-to-T) that has been associated with hereditary persistence of fetal hemoglobin (HPFH). To test for cis-acting determinants located within the introns of the gamma-globin gene, expression was measured from a set of gamma-globin genes configured with either intron alone or with neither intron. In contrast to an intronless beta-globin gene, which is not expressed in MEL cells, the intronless gamma-globin gene was expressed in MEL cells at 24% of the level of an intron-containing gene.(ABSTRACT TRUNCATED AT 250 WORDS)

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mentioning

confidence: 95%

Expression of the human .gamma.-globin gene after retroviral transfer to transformed erythroid cells

Rixon¹,

Harris²,

Gelinas³

1990

Biochemistry

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A high-efficiency vector for expression of foreign genes in myeloma cells

Hendricks

Banker

Mclaughlin

1988

Gene

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Expression of transfected vimentin genes in differentiating murine erythroleukemia cells reveals divergent cis-acting regulation of avian and mammalian vimentin sequences.

Ngai¹,

Bond²,

Wold³

et al. 1987

Mol. Cell. Biol.

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We studied the expression of transfected chicken and hamster vimentin genes in murine erythroleukemia (MEL) cells. MEL cells normally repress the levels of endogenous mouse vimentin mRNA during inducermediated differentiation, resulting in a subsequent loss of vimentin filaments. (8,29,36,77).The generality and diversity of vimentin expression suggest that its regulation is necessarily complex. This complexity is particularly evident from a comparison of avian and mammalian erythropoiesis. The mature avian erythrocyte is a nucleated biconvex ellipsoidally shaped cell. Ultrastructural analyses of avian erythrocytes have revealed a network of intermediate filaments that spans the cytoplasm and appears to anchor the centrally located nucleus (34,89,92). The major subunit protein of these filaments is vimentin (36). * Corresponding author.In contrast, the anucleate, biconcave disk-shaped mammalian erythrocyte contains no intermediate filaments. Studies of human hematopoiesis in vivo have demonstrated that vimentin is expressed early in erythroid differentiation but is lost in the erythroblastic stages (22).In chicken erythroid cells, vimentin filaments are assembled rapidly and stably from a soluble pool of newly synthesized vimentin (5, 62). The efficiency and rapidity of vimentin assembly in vivo suggest that the extent of vimentin filament formation is determined primarily by the amount of vimentin synthesized. We have shown that the expression of vimentin protein during erythropoiesis is determined primarily by mRNA abundance (10, 69). In chicken embryonic erythropoiesis, vimentin mRNA is found at low levels in immature, mitotic primitive cells and accumulates to increasingly higher levels during terminal differentiation of the definitive erythroid lineage, apparently underlying similar changes at the protein level (10). During differentiation of murine erythroleukemia (MEL) cells in vitro, vimentin mRNA levels rapidly and extensively decline (-25-fold reduction), rendering a concomitant decrease in vimentin synthesis and a subsequent loss of vimentin filaments (69). The striking difference in the changes of vimentin mRNA abundances (and, ultimately, vimentin filaments) during mammalian and avian erythropoiesis presents an opportunity to study not only the factors responsible for the dynamic positive and negative regulation of the vimentin gene but also the basis for the evolutionary divergence of the two erythropoietic programs with regard to vimentin expression.In the present study, we addressed these issues by examining the behavior of transfected chicken and hamster vimentin genes in differentiating MEL cells. The utility of studying both resident and transfected genes during in vitro differentiation of MEL cells is well established (reviewed in references 13 and 55). We demonstrate that, in MEL cell lines harboring and expressing chicken vimentin genes,

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Expression of Transfected Genes

Cited by 4 publications

References 65 publications

Expression of the human .gamma.-globin gene after retroviral transfer to transformed erythroid cells

Expression of the human .gamma.-globin gene after retroviral transfer to transformed erythroid cells

A high-efficiency vector for expression of foreign genes in myeloma cells

Expression of transfected vimentin genes in differentiating murine erythroleukemia cells reveals divergent cis-acting regulation of avian and mammalian vimentin sequences.

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