Full Length Research Paper cDNA Cloning and Expression of ATP Synthase Subunit b from the Fire-bellied Frog Bombina orientalis (Anura: Discoglossidae) MIN-SUN KIM a , IL-CHAN KIM b , SANG-SOO KIM a , SOOK-KYOUNG KIM c , YOUNG-MI LEE c , HAN SEUNG KANG a , HEE-SEOK KWEON c , UI-WOOK HWANG d , MOON KYOO KIM a , JAE-SEONG LEE c, * and YONG-DAL YOON a,† a Department of Life Science, College of Natural Sciences, Hanyang University, Seoul 133-791, South Korea; b Department of Biochemistry, College of Medicine, Hanyang University, Seoul 133-791, South Korea; c Department of Environmental Science, Graduate School, Hanyang University, Seoul 133-791, South Korea; d Department of Biology, Teachers College, Kyungpook National University, Taegu 702-701,South Korea (Received 13 January 2004) We cloned the Bombina orientalis adenosine triphosphate (ATP) synthase subunit b gene from a B. orientalis oviduct cDNA library. The transcript was 997 bp long and encoded 250 amino acid residues. It showed high similarity to amphibian (84 – 85%), mammalian (56 – 62%) and invert- ebrate (46 – 50%) sequences. In phylogenetic analyses, it clustered with other amphibian sequences. This gene was highly expressed in brain, intestine and oviduct but not in muscle and liver. In this paper, we report the basic characteristics of B. orientalis ATP synthase subunit b gene. Keywords: Frog; Bombina orientalis; ATP synthase; cDNA Database Accession Number: AY522571 INTRODUCTION Adenosine triphosphate (ATP) is one of the major energy sources in living organism. F1F0-ATP synthase produces ATP in cells and is able to hydrolyze ATP (Walker et al., 1991; Belogrudov and Hatefi, 2002). This enzyme is consisted of two rotary motors, the F1 part (linked to catalytic site events with movements of an internal rotor) and the F0 part (linked to proton translocation to movements of the F0 rotor). These two parts are composed of several subunits linked by two stalks; one is the central stalk comprised of g and 1 subunits and the other is the peripheral stalk comprised of d and b subunits. F1 consists of a, b, g, d and 1 subunits, while F0 consists of a, b and c subunits. ATP synthase can be divided into several subcomplexes; catalytic site, central stalk and peripheral stalk. Of these subunits, the a 3 b 3 acts as a catalytic site and ATP catalytic activity is obtained by conformational change of b subunit. During ATP synthesis, a 3 b 3 subunit rotation causes rotation of the g1c ring , central stalk. This rotation of g1c ring relative to subunit a is critical for proton transport. Also subunit b and d form a stator, preventing a 3 b 3 from rotating with g1, so that the conformational changes can be effected (Weber and Senior, 2003). Biochemically, subunit b of the F0 part of Escherichia coli ATP synthase, composing peripheral stalk with d subunit, can be divided into four domains, the N-terminal membrane-spanning domain, tether domain, dimerization domain and C-terminal d-binding domain (Dunn et al., 2000). However, there is little information available on the similarity of components of ATP synthesizing enzymes in diverse species. Sequence information on the ATP synthase subunit b gene is available from mammals such as human (Farrell and Nagley, 1987; Higuti et al., 1991; Strausberg et al., 2002), mouse (Kawai et al., 2001), rat (Tsurumi et al., 1990) and cow (Walker et al., 1987) ISSN 1042-5179 print/ISSN 1029-2365 online q 2004 Taylor & Francis Ltd DOI: 10.1080/10425170410001723930 *Corresponding author. Tel.: þ 82-2-2290-0769. Fax: þ 82-2-2299-9450. E-mail: jslee2@hanyang.ac.kr † Tel.: þ 82-2-2290-0955. E-mail: ydyoon@hanyang.ac.kr DNA Sequence, June 2004 Vol. 15 (3), pp. 196–201 Mitochondrial DNA Downloaded from informahealthcare.com by Sung Kyun Kwan University on 04/16/14 For personal use only.