Regulation of Na+,K+-ATPases I. Cloning and analysis of the 5′-flanking region of the rat NKAA2 gene encoding the α2 subunit

Kawakami, Kiyoshi; Yagawa, Yuriko; Nagano, Kei

doi:10.1016/0378-1119(90)90098-c

Cited by 19 publications

(10 citation statements)

References 12 publications

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“…No other Na,K-ATPase subunit genes have sequence elements similar to that of the ARE (16,18,21,34). The active transcription factor (ATF/CREB) consensus binding sequence (22,36) is found from -71 to -65 (GTGACGT), but no binding factor was found in a gel retardation assay using a DNA fragment from -102 to -58 (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…To reveal the molecular mechanisms of the tissue-specific, developmental, and hormonal regulation of Na,K-ATPase gene expression and of the coordinated regulation between the a-and 13-subunit genes, we have performed a systematic analysis of the genes encoding a-and 13-subunit isoforms, including their 5'-flanking regions where important regulatory functions reside (15,17,18,43). We cloned from the rat chromosome a 13.3-kb DNA fragment that included the Na,K-ATPase al subunit gene (ATPlA1) and 5' flanking region (EMBL/GenBank data library accession number X51461), and identified the transcription initiation site at 262 bp upstream from the translation initiation codon (43).…”

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confidence: 99%

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Housekeeping Na,K-ATPase alpha 1 subunit gene promoter is composed of multiple cis elements to which common and cell type-specific factors bind.

Suzuki-Yagawa¹,

Kawakami²,

Nagano³

1992

Mol. Cell. Biol.

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Na,K-ATPase al subunit gene (ATPlA1) is one of the housekeeping genes involved in homeostasis of Na+ and K+ in all animal cells. We identified and characterized the cis-acting elements that regulate the expression of ATPAL1. The region between -155 and -49 was determined as a positive regulatory region in five cultured cell lines of different tissue origins (MDCK, B103, L6, 3Y1, and HepG2). The region was divided into three subregions: from -120 to -106 (including the Spl binding site), from -102 to -61, and from -58 to -49 (including an Spl consensus sequence). Cell type-specific factors binding to the middle subregion (from -102 to -61) were detected by gel retardation analysis, using nuclear extracts prepared from MDCK and B103 cells. Two gel retardation complexes were formed in the B103 nuclear extract, and three were formed in the MDCK nuclear extract. DNA binding regions of these factors were located at -88 to -69 and differed from each other in DNase I footprinting experiments. These factors also showed different binding characteristics in gel retardation competition and methylation interference experiments. The identified cis element was named the ATPL1A regulatory element. The core sequence of this element is found in several other genes involved in cellular energy metabolism, suggesting that the sequence is a common regulatory element responsive to the state of energy metabolism.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Housekeeping Na,K-ATPase alpha 1 subunit gene promoter is composed of multiple cis elements to which common and cell type-specific factors bind.

Suzuki-Yagawa¹,

Kawakami²,

Nagano³

1992

Mol. Cell. Biol.

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“…Cells were incubated at 37°C for 6 h in 95% air and 5% CO2. They were washed twice with PBS containing 137 mM NaCl, 2.7 mM KC1, 1.5 mM KH2PO4, and 8.1 mM Na2HPO4 before being fixed with 1% para-formaldehyde and 0.2% Triton-X for 15 min (31), a 2.5-kb BstE II fiagment of the a2 isoform gene from the 5'-flanking region to just before the translation initiation site in the first exon (+60) (32), and a 2.6-kb Hind III-Sac II fragment of the a3 isoform gene from the 5'-flanking region to just before the translation initiation site in the first exon (+ 139) (33), were prepared for the reporter gene assay. Each fragment was ligated to a Hind III-digested pSVOA/LA5' vector (pa lLF, pa2LF, pa3LF).…”

Section: Methodsmentioning

confidence: 99%

Regulation of Na,K-adenosine triphosphatase gene expression by sodium ions in cultured neonatal rat cardiocytes.

Yamamoto¹,

Ikeda²,

Seino³

et al. 1993

J. Clin. Invest.

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Na,K-ATPase (NaK-pump) plays an important role in the regulation of intracellular ion composition. The purpose of this study is to determine whether Na' regulates the levels of mRNA coding for NaK-ATPase a and fi subunits in cultured neonatal rat cardiocytes. We measured intracellular Na+ levels (QNa'Ij) in cardiocytes using a Na'-sensitive fluorescence dye (SBFI). 1 mM ouabain caused a significant increase in [Nalji in cardiocytes; from 12.8±03 to 28.8±1.8 mM. Exposure of cardiocytes to 1 mM ouabain resulted in a three-to fourfold increase in al, a2, and a3 mRNA accumulation, and an approximate two-fold increase in fB1 mRNA accumulation. A maximum elevation was reached at 60 min in both cases. The ouabain-induced al mRNA accumulation was still observed in the Ca2"-free culture medium. Exposure of cardiocytes to 10 gM monensin in the absence of extracellular Ca2" also resulted in a threefold increase in al mRNA accumulation. The increased al mRNA expression by 1 mM ouabain was associated with a fourfold increase in al subunit protein accumulation. Transfection experiments with chimeric plasmids containing 5'-flanking sequences of al, a2, and a3 isoform genes and a luciferase reporter gene revealed that 1 mM ouabain caused a twofold increase in luciferase activity in each a system. These results suggest that Na+ directly regulates NaK-ATPase gene expression in cardiocytes. The transfection study further supports the premise that Na+-responsive elements are located within the 5'-flanking sequences of each a isoform gene. (J.

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“…The BstEII -BstEtI fragment spanning from nucleotide -2406 of 5'-flanking region (the nucleotides are numbered with reference to the transcription initiation site, designated as +1) to nucleotide +60 was excised from Atpla2 [14]. Both ends were filled in with Klenow fragment and were coupled with XhoI linkers.…”

Section: Plasmid Constructionmentioning

confidence: 99%

Anomalous interaction of Sp1 and specific binding of an E‐box‐binding protein with the regulatory elements of the Na,K‐ATPase α2 subunit gene promoter

Ikeda

Nagano

Kawakami

1993

European Journal of Biochemistry

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We identified cis elements in the 5'-flanking region of rat Na,K-ATPase a2 subunit gene (Atpl a2) using transient transfection assays in L6 rat skeletal muscle myoblast cells. By 5'-deletion mutation analysis, the region between nucleotide positions -175 and -108 was identified as a positive regulatory region. In the region, the distal E box (nucleotides -144 to -139) acts as a negative regulatory element, and the Spl consensus sequence (nucleotides -123 to -118) and the GGGAGG sequence (nucleotides -114 to -109) act as positive regulatory elements. Gel-retardation analysis revealed that binding factors are an E-box-binding protein and Spl. DNase I footprinting and methylation-interference analyses revealed that Spl binds to the region from nucleotides -122 to -101 and the E-box-binding protein to the region from nucleotides -144 to -136. T4 DNA polymerase footprinting revealed that there are three Spl-binding sites in the region and that Spl binds to one of the three sites in a mutually exclusive manner. The mechanism by which Spl activates the Atpl a2 promoter is discussed.

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Regulation of Na+,K+-ATPases I. Cloning and analysis of the 5′-flanking region of the rat NKAA2 gene encoding the α2 subunit

Cited by 19 publications

References 12 publications

Housekeeping Na,K-ATPase alpha 1 subunit gene promoter is composed of multiple cis elements to which common and cell type-specific factors bind.

Housekeeping Na,K-ATPase alpha 1 subunit gene promoter is composed of multiple cis elements to which common and cell type-specific factors bind.

Regulation of Na,K-adenosine triphosphatase gene expression by sodium ions in cultured neonatal rat cardiocytes.

Anomalous interaction of Sp1 and specific binding of an E‐box‐binding protein with the regulatory elements of the Na,K‐ATPase α2 subunit gene promoter

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