Human Globin Gene Promoter Sequences Are Sufficient for Specific Expression of a Hybrid Gene Transfected into Tissue Culture Cells

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 1987 Jan 1

PMID 3561396

Citations 22

Authors

T Rutherford

A W Nienhuis

Affiliations

Soon will be listed here.

Abstract

The contribution of the human globin gene promoters to tissue-specific transcription was studied by using globin promoters to transcribe the neo (G418 resistance) gene. After transfection into different cell types, neo gene expression was assayed by scoring colony formation in the presence of G418. In K562 human erythroleukemia cells, which express fetal and embryonic globin genes but not the adult beta-globin gene, the neo gene was expressed strongly from a fetal gamma- or embryonic zeta-globin gene promoter but only weakly from the beta promoter. In murine erythroleukemia cells which express the endogenous mouse beta genes, the neo gene was strongly expressed from both beta and gamma promoters. In two nonerythroid cell lines, human HeLa cells and mouse 3T3 fibroblasts, the globin gene promoters did not allow neo gene expression. Globin-neo genes were integrated in the erythroleukemia cell genomes mostly as a single copy per cell and were transcribed from the appropriate globin gene cap site. We conclude that globin gene promoter sequences extending from -373 to +48 base pairs (bp) (relative to the cap site) for the beta gene, -385 to +34 bp for the gamma gene, and -555 to +38 bp for the zeta gene are sufficient for tissue-specific and perhaps developmentally specific transcription.

Citing Articles

Positive selection of DNA-protein interactions in mammalian cells through phenotypic coupling with retrovirus production.

Tschulena U, Peterson K, Gonzalez B, Fedosyuk H, Barbas 3rd C Nat Struct Mol Biol. 2009; 16(11):1195-9.

PMID: 19838191 PMC: 2880176. DOI: 10.1038/nsmb.1677.

Identification and characterization of mechanistically distinct inducers of gamma-globin transcription.

Haley J, Smith D, Schwedes J, Brennan R, Pearce C, Moore C Biochem Pharmacol. 2003; 66(9):1755-68.

PMID: 14563486 PMC: 1351252. DOI: 10.1016/s0006-2952(03)00542-2.

Methylation-enhanced binding of Sp1 to the stage selector element of the human gamma-globin gene promoter may regulate development specificity of expression.

Jane S, Gumucio D, Ney P, Cunningham J, Nienhuis A Mol Cell Biol. 1993; 13(6):3272-81.

PMID: 7684493 PMC: 359778. DOI: 10.1128/mcb.13.6.3272-3281.1993.

Gene expression as a target for new drug discovery.

Heguy A, Stewart A, Haley J, Smith D, Foulkes J Gene Expr. 1995; 4(6):337-44.

PMID: 7549465 PMC: 6134365.

Activation of the human beta-globin promoter in K562 cells by DNA sequences 5' to the fetal gamma- or embryonic zeta-globin genes.

Lin H, Anagnou N, Rutherford T, Shimada T, Nienhuis A J Clin Invest. 1987; 80(2):374-80.

PMID: 3611352 PMC: 442247. DOI: 10.1172/JCI113082.

References

Southern E . Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975; 98(3):503-17. DOI: 10.1016/s0022-2836(75)80083-0. View

Myers R, Tilly K, Maniatis T . Fine structure genetic analysis of a beta-globin promoter. Science. 1986; 232(4750):613-8. DOI: 10.1126/science.3457470. View

Rutherford T, Clegg J, Weatherall D . K562 human leukaemic cells synthesise embryonic haemoglobin in response to haemin. Nature. 1979; 280(5718):164-5. DOI: 10.1038/280164a0. View

Dierks P, Ooyen A, Mantei N, Weissmann C . DNA sequences preceding the rabbit beta-globin gene are required for formation in mouse L cells of beta-globin RNA with the correct 5' terminus. Proc Natl Acad Sci U S A. 1981; 78(3):1411-5. PMC: 319140. DOI: 10.1073/pnas.78.3.1411. View

Rutherford T, Clegg J, Higgs D, Jones R, Thompson J, Weatherall D . Embryonic erythroid differentiation in the human leukemic cell line K562. Proc Natl Acad Sci U S A. 1981; 78(1):348-52. PMC: 319050. DOI: 10.1073/pnas.78.1.348. View

Grosveld G, Shewmaker C, Jat P, Flavell R . Localization of DNA sequences necessary for transcription of the rabbit beta-globin gene in vitro. Cell. 1981; 25(1):215-26. DOI: 10.1016/0092-8674(81)90246-4. View

Grosveld G, de Boer E, Shewmaker C, Flavell R . DNA sequences necessary for transcription of the rabbit beta-globin gene in vivo. Nature. 1982; 295(5845):120-6. DOI: 10.1038/295120a0. View

Southern P, Berg P . Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982; 1(4):327-41. View

Ley T, Anagnou N, Pepe G, Nienhuis A . RNA processing errors in patients with beta-thalassemia. Proc Natl Acad Sci U S A. 1982; 79(15):4775-9. PMC: 346760. DOI: 10.1073/pnas.79.15.4775. View

10.

Grosveld G, Rosenthal A, Flavell R . Sequence requirements for the transcription of the rabbit beta-globin gene in vivo: the -80 region. Nucleic Acids Res. 1982; 10(16):4951-71. PMC: 320844. DOI: 10.1093/nar/10.16.4951. View

11.

Gorman C, Moffat L, Howard B . Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982; 2(9):1044-51. PMC: 369897. DOI: 10.1128/mcb.2.9.1044-1051.1982. View

12.

Chao M, Mellon P, Charnay P, Maniatis T, Axel R . The regulated expression of beta-globin genes introduced into mouse erythroleukemia cells. Cell. 1983; 32(2):483-93. DOI: 10.1016/0092-8674(83)90468-3. View

13.

Dierks P, Ooyen A, Cochran M, Dobkin C, Reiser J, Weissmann C . Three regions upstream from the cap site are required for efficient and accurate transcription of the rabbit beta-globin gene in mouse 3T6 cells. Cell. 1983; 32(3):695-706. DOI: 10.1016/0092-8674(83)90055-7. View

14.

Crouse G, Frischauf A, Lehrach H . An integrated and simplified approach to cloning into plasmids and single-stranded phages. Methods Enzymol. 1983; 101:78-89. DOI: 10.1016/0076-6879(83)01006-x. View

15.

Wright S, DEBOER E, Grosveld F, Flavell R . Regulated expression of the human beta-globin gene family in murine erythroleukaemia cells. Nature. 1983; 305(5932):333-6. DOI: 10.1038/305333a0. View

16.

Spandidos D . Expression of the human beta globin gene and 5'-deletion mutants in erythroleukemic mouse cells studied by DNA mediated gene transfer. Anticancer Res. 1983; 3(6):409-16. View

17.

Neumann E, Wang Y, Hofschneider P . Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J. 1982; 1(7):841-5. PMC: 553119. DOI: 10.1002/j.1460-2075.1982.tb01257.x. View

18.

Episkopou V, Murphy A, Efstratiadis A . Cell-specified expression of a selectable hybrid gene. Proc Natl Acad Sci U S A. 1984; 81(15):4657-61. PMC: 391549. DOI: 10.1073/pnas.81.15.4657. View

19.

Collins F, Stoeckert Jr C, Serjeant G, Forget B, Weissman S . G gamma beta+ hereditary persistence of fetal hemoglobin: cosmid cloning and identification of a specific mutation 5' to the G gamma gene. Proc Natl Acad Sci U S A. 1984; 81(15):4894-8. PMC: 391598. DOI: 10.1073/pnas.81.15.4894. View

20.

Charnay P, Treisman R, Mellon P, Chao M, Axel R, Maniatis T . Differences in human alpha- and beta-globin gene expression in mouse erythroleukemia cells: the role of intragenic sequences. Cell. 1984; 38(1):251-63. DOI: 10.1016/0092-8674(84)90547-6. View