Structure and organization of the rat Endothelin-2 gene

Structure and organization of the rat Endothelin-2 gene Brief Data Reports Mammalian Genome 8, 157-160 (1997). 9 Springer-Verlag New York Inc. 1997 For Edn2 to be considered as a valid candidate gene for being the Structure and organization of the rat blood pressure QTL on rat Chr 5, there would have to be a sig- Endothelin-2 gene nificant nucleotide difference(s) between the Edn2 gene of the S rat and the Edn2 gene of the LEW rat. This difference(s) would be Alan Y. Deng such that it could cause the differing Edn2 genes of the S and LEW rats to function differently. Department of Physiology and Molecular Medicine, Block Health In the current work, there are no significant nucleotide differ- Science Building, Room 320, Me&cal College of Ohio, P.O. Box ences between the S and LEW rats detected in the region of Edn2 10008, Toledo, Ohio 43699-0008, USA genes that are potentially important in the Edn2 function. As a consequence, the Edn2 gene per se is not supported as a candidate Received: 5 June 1996 / Accepted: 4 October 1996 QTL causing a blood pressure difference between the S and LEW rats [1]. The present sequence data alone, however, do not, and Species: Rat (Rattus norvegicus) cannot, rule Edn2 out as the blood pressure QTL, because uniden- Locus name: Endothelin-2 tified regulatory regions either located in the 5' or 3' end or in Locus symbol: Edn2 intronic regions of Edn2 could still be potentially important in Map position: Edn2 is located on rat Chromosome (Chr) 5 [1]. differentially regulating the Edn2 gene expression between the two Database deposit information: GenBank Accession Numbers: rat strains. To definitely exclude Edn2 as the blood pressure QTL, U59510, U64946--U64949. one has to show that the chromosome fragment containing Edn2 Molecular reagents: Sequencing was done using a Sequenase Kit from the LEW rat, after replacing the homologous chromosome (Amersham, Arlington Heights, Ill.) region of the S rat, would not significantly lower blood pressure in Cloning ofEdn2: Genomic clones containing Edn2 were obtained a congenic rat. by screening a rat genomic DNA library with a 129 base-pair Edn2 probe [1]. The genomic organization of Edn2 is shown in Fig. 1. Acknowledgment. I thank Dr. John Rapp for providing rat DNA samples, Sequence comparison of Edn2: Edn2 genomic fragments contain- Dr. George Cicila for help in analyzing the 5' flanking region of Edn2 by ing the 5' flanking region, all exons including the 3' untranslated computer, and Stephanie Mirarck for technical assistance. This work was region and all the exon-intron junctions, were PCR amplified from supported by a Grant-in-Aid from the American Heart Association and DNA of both the inbred hypertensive Dahl salt-sensitive (S) rat attributed in part by the American Heart Association, Ohio Affiliate. strain and the Lewis (LEW) rat. These sequences were compared. References So far, no nucleotide differences were found between the Edn2 1. Deng AY, Dene H, Pravenec M, Rapp JP (1994) J Clin Invest 93, genes of the two rat strains. 2701-2709 Discussion: A quantitative trait locus (QTL) for blood pressure 2. Rapp JP, Deng AY (1995) Hypertension 25, 1121-1128 has been localized to a region of rat Chr 5 in the S rat [1]. The 3. Inoue A, Yanagisawa M, Kimura S, Kasuya Y, Miyanchi T, Goto K, chromosome region harboring the QTL contains several genes that Masakl T (1989) Proc Natl Acad Sci USA 86, 28363-2867 can be construed as logical candidates as the QTL because they are potentially important in blood pressure regulation [2]. Of these candidate genes closest to the predicted position of the QTL on The Tage4 gene maps to rat Chr, the Edn2 gene is obvious since endothelin-2 peptide was Chromosome lq22 found to increase blood pressure in rats after a bolus injection [3]. C. Chad6neau, 1 T. Liehr, 2 B. Rautenstrauss, 2 M.G. Denis 1 8 X X B I I I I IINSERM U419, Institut de Biologie, 9 Quai Moncousu, F-44035 Nantes, France 2Institute of Human Genetics, Schwabachanlage 10, D-91054 Erlangen, Germany IV Received: 15 July 1996 / Accepted: 25 September 1996 5" '~ 3' (1] Species: Rat (Rattus norvegicus) Locus name: Tumor-associated glycoprotein pE4 t~ Locus symbol: Tage4 Map position: Chromosome (Chr) lq22 Method of mapping: Fluorescence in situ hybridization (FISH) as described [ 1,2], performed on chromosomes derived from the male Fig. 1. Cloning and physical mapping of the rat Edn2 gene. (a) A genomic fibroblast cell line RAT1. fragment acquired from screening a rat genomic library with an Edn2 Molecular reagents used for mapping: KS-pE4.1 contains a 1423- specific probe. B, BamHl and X, XbaI resmction enzymes. (b) Physical bp eDNA insert corresponding to the coding region of the rat mapping of Edn2. Black blocks I to V represent exons. The upward- Tage4 eDNA [3], cloned in the EcoRI site of the pBluescript KS pointing arrow at exon V indicates the posltion of the stop codon. (c) Line 1 represents the Xbal fragment in (a); how far it extends towards the 5' and plasmid. 3' ends of the Edn2 gene has not been determined (dots symbolize the ambiguity). Lines 2 to 6 denote overlapping PCR-amplified genomic frag- Correspondence to." M.G. Denis ments that cover most of the Edn2 gene. Mammalian Genome Springer Journals

Structure and organization of the rat Endothelin-2 gene

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