Nucleic Acids Research, Vol. 18, No. 16 4923
.=) 1990 Oxford University Press
Nucleotide sequence of cDNA for the cytosolic Cu/Znsuperoxide dismutase from spinach (Spinacia oleracea L.) Atsushi Sakamoto, Hiroyuki Ohsuga, Makoto Wakaura, Norihiro Mitsukawa, Takashi Hibino, Takehiro Masumura, Yukiko Sasaki' and Kunisuke Tanaka Department of Biochemistry, College of Agriculture, Kyoto Prefectural University, Shimogamo, Kyoto 606 and 'Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Kyoto 606, Japan Submitted July 14, 1990
EMBL accession
Superoxide dismutase (SOD; EC 1.15.1.1),
a
metalloenzyme
Supported in part by a Grant from the Ministry of Agriculture, Forestry and Fisheries of Japan and also by a Grant 63860004 from the Ministry of Education, Science and Culture of Japan.
defence against oxygen toxicity by catalyzing the disproportionation of superoxide anion radicals. Three types of SOD (Cu/Zn-, Mn- and Fe-containing enzymes) have been found among the eukaryotes. Cu/Zn-SOD is the major type found in higher plants, and is classified into two types: that in the cytosol and the other in the chloroplast (1, 2). Using a 75-mer synthetic oligonucleotide probe complementary to the cDNA sequence of maize cytosolic SOD (3), a cDNA clone encoding the cytosolic Cu/Zn-SOD of spinach was isolated from the Xgtl library derived from the leaf mRNA. Its identity was confirmed by comparison of the sequence of the clone to the NH2- and COOH-terminal sequences of the protein (4). The deduced sequence of 152 amino acids showed higher homology with the sequences of the cytosolic SOD from several other plants (maize, 79%; tomato, 84%) (3, 5) than the sequence of the chloroplastic counterpart of spinach (69%) (6). Seven residues coordinating copper and zinc were conserved as reported for the Cu/Zn-SOD
REFERENCES 1. Asada,K. (1988) In Otsuka,S. and Yamanaka,T. (eds) Metalloproteins, Elsevier, Amsterdam, pp. 331-341. 2. Asada,K., Kanematsu,S., Okada,S. and Hayakawa,T. (1980) In Bannister,J.V. and Hill,H.A.O. (eds), Chemical and Biochemical Aspects of Superoxide and Superoxide Dismutase, Elsevier, Amsterdam, pp. 136- 153. 3. Cannon,R.E., White,J.A. and Scandalios,J.G. (1987) Proc. Natl. Acad. Sci. USA 84, 179-183. 4. Kanematsu,S. and Asada,K. (1990) Plant Cell Physiol. 31, 99-112. 5. Perl-Treves,R., Nacmias,B., Aviv,D., Zeelon,E.P. and Galun,E. (1988) Plant Mol. Biol. 11, 609-623. 6. Kitagawa,Y., Tsunasawa,S., Tanaka,N., Katsube,Y., Sakiyama,F. and Asada,K. (1986) J. Biochem. 99, 1289-1298.
sequences. CTCGAATCTTCAACTCCTCTCTCTTTCTCTCTCCTCCAAATTCCAAGGGTGATCTGAGAATACACACAAC
70
ATGGGAAAGGCTGTGGTTGTTCTAAGCAGCAGTGAGGGTGTTTCTGGAACAGTCTACTTTGCCCAGGAAGGA M G K A V V V L S S S E G V S G T V Y F A Q E G
142
I
GATGGTCCAACAACTGTTACTGGAAATGTCTCCGGTCTCAAGCCTGGGCTCCATGGATTTCATGTTCATGCC D G P T T V T G N V S G L K P G L H G F H V H A
214
25
CTTGGTGACACAACAAATGGCTGCATGTCAACTGGGCCGCACTATAACCCAAATGGAAAAGAGCATGGTGCT
286
L
G
D
T
T
N
G
C
M
S
T
X53872
ACKNOWLEDGEMENTS
present in all aerobic organisms, plays an important role in
49
no.
G
P
H
Y
N
P
N
G
K
E
H
G
A
CCAGAAGACGATGTTCGCCATGCTGGTGATCTTGGAAACATCACAGTTGGGGATGATGGTACTGCTACCTTT P E D D V R H A G D L G N I T V G D D G T A T F
358
73
ACCATCATTGACAGTCAGATACCTCTTTCTGGACCCAACTCTATTGTGGGGAGGGCCGTTGTTGTCCATGCT T I I D S Q I P L S G P N S I V G R A V V V H A
430
97
GAACCCGATGATCTTGGAAGGGGTGGACATGAACTCAGCAAGACCACAGGCAATGCTGGTGGAAGAGTGGCT E P D D L G R G G H E L S K T T G N A G G R V A
502
121
TGTGGCATTATTGGTCTTCAAGGTTAAGTTGTGCTTTCCAACTTTCAAAGAGAGAAATCGAAGTGCAGATTA C G I I G L Q G
574
145
CAGTTTTCTTGCTATCCATCCAAGATTAGTGTAGGGGTTAAAAAAAAAGGCAAACAAAAAAAGAAGGGATAA
646
CCCCTAAATAAAAATCCAAGTTATTGTGAATTTGTGACTTGAATTGATTATGTAATCCACCAGAGTCTGTTA
718
TATGTACGGATTTTGCAGTGGCTGTGAACCTTGTTGATTTAAATTTCCTGGCTCTATTTGATGGCTTTTGCA
790
ACTTT
795