Nucleic Acids Research, 1992, Vol. 20, No. 21 5669-5676
© 1992
MOLECULAR BIOLOGY |
In vitro and in vivo protein - DNA interactions on the rat erythroid-specific L' pyruvate kinase gene promoter
ICGM, Laboratoire de Recherches en Génétique et Pathologie Moléculaires INSERM U 129, CHU Cochin, 24 rue de Faubourg, Saint Jacques, 75014 Paris, France
Received July 28, 1992. Revised October 6, 1992. Accepted October 6, 1992.
The rat L-type pyruvate kinase gene possesses two alternative tissue-specific promoters, located 472 bp apart; the upstream L' promoter is erythroid-specific and the downstream L' promoter is hepatocyte-specific. The erythroid-specific L' promoter is strongly active in fetal liver at day 17 of gestation, while its activity rapidly decreases thereafter. A L' promoter fragment spanning from nucleotide –320 to +10 with respect to the cap site is able to direct a weak but erythroid-specific transcription in a cell-free system. We have used DNAse I footprinting and gel mobility shift assays to characterize and identify the binding of nuclear factors from both 17-day-old fetal liver and adult liver nuclear extracts to a 320 bp fragment of the 5' flanking region of the gene in vitro. Two clusters of erythroid-specific interactions were found. The proximal cluster consists of two GATA-1 binding sites at –50 bp and –65 bp from the transcription initiation site, immediately downstream of a CACC motif and two G/C-rich elements. The distal cluster of cis-elements, located 130 bp upstream, corresponds to two GATA-1 sequences. These two sequences overlap NF1 motifs interacting with ubiquitous NF1 transcriptional factors in presence of adult hepatic extracts. Furthermore, we have examined in vivo protein-DNA interactions by DMS footprinting in livers of 17-day-old rat fetuses and adult rats. We found that the sites characterized in vitro are occupied in vivo. Therefore, in adult liver the L' promoter, although inactive, nevertheless interacts with ubiquitous factors.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  E. Zanotto, Z. H. Shah, and H. T. Jacobs The bidirectional promoter of two genes for the mitochondrial translational apparatus in mouse is regulated by an array of CCAAT boxes interacting with the transcription factor NF-Y Nucleic Acids Res., January 28, 2007; 35(2): 664 - 677. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. van Wijk, W. W. van Solinge, C. Nerlov, E. Beutler, T. Gelbart, G. Rijksen, and F. C. Nielsen Disruption of a novel regulatory element in the erythroid-specific promoter of the human PKLR gene causes severe pyruvate kinase deficiency Blood, February 15, 2003; 101(4): 1596 - 1602. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Shimizu, Y. Matsuoka, Y. Shinohara, Y. Yoneda, and Y. Tsujimoto Essential Role of Voltage-dependent Anion Channel in Various Forms of Apoptosis in Mammalian Cells J. Cell Biol., January 16, 2001; 152(2): 237 - 250. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Murakami and S. Piomelli Identification of the cDNA for Human Red Blood Cell-Specific Hexokinase Isozyme Blood, February 1, 1997; 89(3): 762 - 766. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Porcher, C. Picat, D. Daegelen, C. Beaumont, and B. Grandchamp Functional Analysis of DNase-I Hypersensitive Sites at the Mouse Porphobilinogen Deaminase Gene Locus J. Biol. Chem., July 21, 1995; 270(29): 17368 - 17374. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Lacronique, S. Lopez, L. Miquerol, A. Porteu, A. Kahn, and M. Raymondjean Identification and Functional Characterization of an Erythroid-specific Enhancer in the L-type Pyruvate Kinase Gene J. Biol. Chem., June 23, 1995; 270(25): 14989 - 14997. [Abstract] [Full Text] [PDF]
|
|