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

Zn-superoxide dismutase from spinach (Spinacia oleracea L.).

Nucleic Acids Research, Vol. 18, No. 16 4923 .=) 1990 Oxford University Press Nucleotide sequence of cDNA for the cytosolic Cu/Znsuperoxide dismutas...
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