Please use this identifier to cite or link to this item: http://hdl.handle.net/10553/49987
Title: Analysis of ferrochelatase expression during hematopoietic development of embryonic stem cells
Authors: Magness, Scott T.
Tugores, Antonio 
Brenner, David A.
UNESCO Clasification: 32 Ciencias médicas
320504 Hematología
Keywords: Ferrochelatase
Hematopoiesis
Stem cells
Issue Date: 2000
Journal: Blood 
Abstract: 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 cognatetrans-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.
URI: http://hdl.handle.net/10553/49987
ISSN: 0006-4971
DOI: 10.1182/blood.V95.11.3568
Source: Blood[ISSN 0006-4971],v. 95, p. 3568-3577
Appears in Collections:Artículos
Show full item record

SCOPUSTM   
Citations

22
checked on Jun 26, 2022

Page view(s)

17
checked on Jun 25, 2022

Google ScholarTM

Check

Altmetric


Share



Export metadata



Items in accedaCRIS are protected by copyright, with all rights reserved, unless otherwise indicated.