|
|
 Previous Article | Table of Contents | Next Article 
Blood, Vol. 95 No. 11 (June 1), 2000:
pp. 3568-3577
Analysis of ferrochelatase expression during hematopoietic
development of embryonic stem cells
Scott T. Magness,
Antonio Tugores, and
David A. Brenner
From the Departments of Medicine and Biochemistry and
Biophysics, Curriculum in Genetics and Molecular Biology, University of
North Carolina at Chapel Hill.
Ferrochelatase, the last enzyme in the heme pathway, chelates
protoporphyrin IX and iron to form heme and is mutated in
protoporphyria. The ferrochelatase gene is expressed in all tissues at
low levels to provide heme for essential heme-containing proteins and
is up-regulated during erythropoiesis for the synthesis of hemoglobin. The human ferrochelatase promoter contains 2 Sp1 cis-elements and GATA and NF-E2 sites, all of which bind their cognate
trans-acting factors in vitro. To investigate the role of these
elements during erythropoiesis, we introduced expression of the green
fluorescent protein (EGFP) transgenes driven by various ferrochelatase
promoter fragments into a single locus in mouse embryonic stem cells.
EGFP expression was monitored during hematopoietic differentiation in
vitro using flow cytometry. We show that a promoter fragment containing
the Sp1 sites, the NF-E2 and GATA elements, was sufficient to confer
developmental-specific expression of the EGFP transgene, with an
expression profile identical to that of the endogenous gene. In this
system the  0.275 kb NF-E2 cis-element is required for
erythroid-enhanced expression, the GATA cis-element functions as a stage-specific repressor and enhancer, and elements located between 0.375kb and 1.1kb are necessary for optimal levels of expression. Ferrochelatase mRNA increased before the primitive erythroid-cell stage without a concomitant increase in ferrochelatase protein, suggesting the presence of a translational control mechanism. Because of the sensitivity of this system, we were able to assess the
effect of an A-to-G polymorphism identified in the promoters of
patients with protoporphyria. There was no effect of the G haplotype on
transcriptional activity of the 1.1 kb transgene.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
W. Kemmner, K. Wan, S. Ruttinger, B. Ebert, R. Macdonald, U. Klamm, and K. T. Moesta
Silencing of human ferrochelatase causes abundant protoporphyrin-IX accumulation in colon cancer
FASEB J,
February 1, 2008;
22(2):
500 - 509.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
I. Puga, B. Lainez, J. M. Fernandez-Real, M. Buxade, M. Broch, J. Vendrell, and E. Espel
A Polymorphism in the 3' Untranslated Region of the Gene for Tumor Necrosis Factor Receptor 2 Modulates Reporter Gene Expression
Endocrinology,
May 1, 2005;
146(5):
2210 - 2220.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. T. Magness, H. Jijon, N. Van Houten Fisher, N. E. Sharpless, D. A. Brenner, and C. Jobin
In Vivo Pattern of Lipopolysaccharide and Anti-CD3-Induced NF-{kappa}B Activation Using a Novel Gene-Targeted Enhanced GFP Reporter Gene Mouse
J. Immunol.,
August 1, 2004;
173(3):
1561 - 1570.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. Zheng, Z. Zhang, C. M. Black, B. de Crombrugghe, and C. P. Denton
Ligand-Dependent Genetic Recombination in Fibroblasts : A Potentially Powerful Technique for Investigating Gene Function in Fibrosis
Am. J. Pathol.,
May 1, 2002;
160(5):
1609 - 1617.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A Veraksa, N McGinnis, X Li, J Mohler, and W McGinnis
Cap 'n' collar B cooperates with a small Maf subunit to specify pharyngeal development and suppress deformed homeotic function in the Drosophila head
Development,
January 9, 2000;
127(18):
4023 - 4037.
[Abstract]
[PDF]
|
|
|
|
|
