Open AccessStructure of the 5′‐Flanking Regulatory Region of the Mouse Gene Encoding the Clearance Receptor for Atrial Natriuretic Peptide
Author(s) -
Yanaka Noriyuki,
Kotera Jun,
Taguchi Ikuhiko,
Sugiura Masaki,
Kawashima Keisuke,
Omori Kenji
Publication year - 1996
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1996.0025n.x
Subject(s) - tata box , biology , microbiology and biotechnology , gene , caat box , regulatory sequence , atrial natriuretic peptide , primer extension , complementary dna , transcription (linguistics) , promoter , start codon , response element , 5' flanking region , transcriptional regulation , genomic dna , genetics , gene expression , messenger rna , linguistics , philosophy , endocrinology
A full‐length cDNA, encoding the mouse atrial natriuretic peptide clearance receptor (ANP‐CR), was isolated from a mouse lung cDNA library. The deduced amino acid sequence of the mouse ANP‐CR, showing a typical tripartite organization which lacks a guanylyl cyclase domain, was extremely well conserved compared with the ANP‐CR homologs. To understand the molecular mechanisms underlying the regulation of mouse ANP‐CR gene expression and to define the essential DNA sequences for the transcriptional activity, a genomic clone containing over 9 kb of the 5′‐flanking region of the mouse ANP‐CR gene has been isolated from a mouse genomic library. Sequence analysis revealed that the 2.3‐kb region upstream from an ATG codon of the mouse ANP‐CR gene contained a number of putative regulatory elements; TATA box, CAAT box, cAMP response element, AP‐1 and two shear stress responsive elements. Additionally, an unusual feature was the presence of the tandem‐repeated AP‐2‐like elements, which were closely overlapped with SP‐1 element. Promoter analysis using deletion plasmids in mouse Balb/3T3 cells, highly producing ANP‐CR mRNA, demonstrated that deletion of the sequence from −144 to +46 relative to the transcription start point caused a dramatic decrease of the transcriptional activity and that the TATA box at −269 was not essential for the basal transcriptional activity. Primer extension analysis indicated that transcription of the mouse ANP‐CR gene starts from at least two major sites, suggesting that the sequence from −144 to +46, which was shown to involve a novel sequence composed of tandem‐repeated TATA‐box‐like elements, contained promoter sequences. Furthermore, cis acting negative elements were shown to be situated in three regions (from −1178 to −708, from −707 to −625 and from −248 to −145) of the mouse ANP‐CR gene promoter.