cDNA cloning of Na+, K(+)-ATPase alpha-subunit from embryos of the sea urchin, Hemicentrotus pulcherrimus.
Na+, K(+)-ATPase alpha-subunit cDNA of the sea urchin, Hemicentrotus pulcherrimus, was obtained by twice screening prism and gastrula lambda gt10 cDNA libraries using an oligonucleotide probe derived from a mostly conserved region, FSBA (5'-p-(fluorosulfonyl)-benzoyladenosine) binding site of cation transport ATPases. The 5'-end of the non-coding region was determined by primer extension and the region was amplified by 5'-RACE method. The sea urchin alpha-subunit cDNA consists of 4401 nucleotides and encodes 1038 amino acid residues (MW, 114 kDa). The predicted primary structure, except N-terminal region, has similar degree of high homology to various metazoan Na+, K(+)-ATPase alpha-subunits. Alignment of amino acid sequence and a hydropathy profile also predicts eight putative transmembrane segments at least. The phylogenetic tree suspected from alignment of amino acid sequences of 21 species suggests that sea urchin and vertebrate Na+, K(+)-ATPase alpha-subunits seem to have evolved from a common origin, before vertebrate alpha-subunit divided into three isoforms.
Mitsunaga-Nakatsubo
K
K
Department of Biology, School of Education, Waseda University, Tokyo, Japan.
Yamazaki
K
K
Hatoh-Okazaki
M
M
Kawashita
H
H
Okamura
C
C
Akasaka
K
K
Shimada
H
H
Yasumasu
I
I
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004625:Embryo, Nonmammalian / Q000201:enzymology*; D008969:Molecular Sequence Data; D010802:Phylogeny; D012617:Sea Urchins / Q000196:embryology / Q000201:enzymology / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics*
D018076:DNA, Complementary; D000254:Sodium-Potassium-Exchanging ATPase