Mapping of the rat and mouse nicotinamide nucleotide transhydrogenase gene

Mapping of the rat and mouse nicotinamide nucleotide transhydrogenase gene Mammalian Genome 8, Brief Data Reports 703 ing on tail cuff systolic blood pressure and mean intraarterial blood pressure in the male F 2 population of a GH x BN cross [4]. The map location for KBP determined in the present study and the failure to detect any cosegregation with either blood pressure phe- notype in our GH x BN cross allow us to conclude that KBP is not the affected gene at the hypertensive locus we have found on rat Chr 6. References 1. Chai KX, Ma J-X, Murray SR, Chao J, Chao L (1991) J Biol Chem 266, 16029-16036 2. Harris EL, Phelan EL, Millar JA, Grigor MR (1995) J Hypertens 13, 397-404 3. Chomczynski P, Sacchi N (1987) Anal Biochem 162, 156-159 4. Jacob Lab, Cowley Lab, Dzau Lab, Grigor Lab, Fanesyl Lab, Harrap Lab, Harris Lab, Krieger EM Lab, Krieger JE Lab, Lander Lab, Prinz Lab, Provoost Lab, Roman Lab, Sassard Lab, Schork Lab, Vincent Lab (1996) J Hypertens 14 (Suppl 1), $7 5. Chao J, Chao L (1995) Biol Chem Hoppe Seyler 376, 705-713 6. Chao J, Chao L (1988) J Hypertens 6, 551-557 7. Yang T, Terada Y, Nonoguchi H, Tsujino M, Tomita K, Marumo F (1994) Am J Physiol 267, F325-F330 8. Lindpaintner K, Chai K, Kreutz R, Hubner N, Ganten D, Chao L, Chao Fig. 1. Mapping of the mouse and rat Nnt genes by FISH. a, c. Hybrid- J (1994) Clin Exp Pharmacol Physiol 20(1), $3 ization signals (arrow-heads) at MMU13D2 and RNO2q16, respectively, b, d. Same metaphase areas in 4,6/diamino/2/phenylindole staining. The col- ors have been artificially reversed to reveal a G-bandlike pattern. Mapping of the rat and mouse nicotinamide nucleotide transhydrogenase gene for EST01744 (locus designation D5S523E; [5]). The EST01744 is 264bp long, and we found that 262 of the bp were identical to a Eva Lagberg Arkblad, 1 Khalil Helou, 2 Gbran Levan, 2 segment of our human NNT cDNA sequence near the central part Jan Rydstr6m 1 (positions 1791 to 2054 from the translation start). The remaining 1Department of Biochemistry & Biophysics, Chalmers University of two bases of EST01744 were both designated as "unknown" in Technology and Grteborg University, S-413 90 Grteborg, Sweden the databank sequence and correspond to "G" in our sequence. 2Department of Genetics, Gtiteborg University, S-413 90 Subsequently, we mapped the mouse and rat genes using the G/3teborg, Sweden mouse NNT cDNA as the probe. The Nnt gene was localized to mouse Chr (MMU) 13D2 (Fig. la,b) and to rat Chr (RNO) 2q16 Received: 19 February 1997 / Accepted: 1 May 1997 (Fig. lc,d). The human NNT gene is included in a group of seven Species: Rat and mouse genes (other members: DHFR, HTRA1, HMGCS1, PDE4D, Locus name: Nicotinamide nucleotide transhydrogenase; MLV12, PRLR) in the region 5p14-q13 (spanning the centro- NAD(P)+ transhydrogenase, (B-specific) mere), which exhibits conserved synteny on RNO2. Most genes on Locus symbol: Nnt RNO2 have homologs on MMU3, but among the genes mentioned, Map position: Rat: Chr 2, band ql6; Mouse: Chr 13, band D2 five genes map on MMU13, and the remaining two on MMU15. Method of mapping: Fluorescence in situ hybridization (FISH), Thus, the NNT gene fits into a group of genes that exhibits con- essentially according to [1] with minor modifications [2]. served synteny between HSA5 (centromeric region), RNO2, and Molecular reagents: Mouse cDNA clone 15 containing the whole MMU13. coding region of the Nnt gene and a 500-bp 3' untranslated region (vector pBluescript). Human cDNA clones H1,H2, and H3, to- Acknowledgments: This work was supported by the Swedish Cancer So- gether containing the whole coding region of the NNT gene and a ciety, Swedish Natural Sciences Research Council, the Trygger Founda- total of 300 bp in the 5' and 3' untranslated regions (vector pBlue- tion, the IngaBritt and Ame Lundberg Research Foundation, and the Erik script) [3]. Philip-Srrensen Foundation. K. Helou is the recipient of a Research Sti- Previously identified homologs: The human D5S523 locus char- pend from the Sven and Lilly Lawski Foundation. acterized by EST01744 (see below) is at HSA5 [4,5]. Discussion: Eukaryotic transhydrogenases are membrane proteins and proton pumps that generate NADPH at the expense of NADH References and are located in the mitochondrial inner membrane. The physi- 1. Pinkel D, Landegent J, Collins C, Fuscose J, Segraves R, Lucas J, Gray ological role of NAD(P)+ transhydrogenase has not been fully J (1988) Proc Natl Acad Sci USA 85, 9138-9142 established, but has been proposed to involve redox regulation of metabolism and proteins, and provide defense against free radicals 2. Szpirer C, Tissir F, Rivi&e M, Levan G, Szpirer J (1994) Mamm [6]. Human and mouse NNT cDNAs were isolated and completely Genome 5, 361-364 sequenced as described (GenBank Accession No. Z50101 and 3. Lagberg Arkblad E, Betsholtz C, Rydstrrm J (1996) Biochim Biophys Z49204, respectively [3]). Searching the GenBank database, we Acta 1273, 203-205 came across a human EST01744 (Accession No. M78152), de- 4. Durkin AS, Nierman WC, Zoghbi H, Jones C, Kozak CA, Maglott DR scribed as "similar to NAD(P)+ transhydrogenase, B-specific". (1994) Cytogenet Cell Genet 65, 86--91 Our human NNT probe mapped to human Chr (HSA) 5pl3.1--cen 5. Feldblyum TV, Maglott DG, McPherson JD, Adams M, Apostol BL, (data not shown), which is the same localization as that reported Durkin AS, Wasmuth JJ, Nierman WC (1996) Genomics 33, 128-130 6. Olausson T, Fjellstrrm O, Meuller J, Rydstr0m J (1995) Biochim Bio- phys Acta 1231, 1-19 Correspondence to: E. Lagberg Arkblad Mammalian Genome Springer Journals

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