Cloning and characterization of a cDNA encoding a rice 13 kDa prolamin.
A cDNA library constructed from mRNAs obtained from developing rice endosperm was screened with a cDNA clone (lambda RM7) of highest frequency of occurrence (1.8%). The translation product directed by the mRNA which was hybrid-released from lambda RM7 cDNA in a wheat germ cell-free system showed a molecular size of 13 kDa when coexisting with the protein body fraction of developing maize endosperm. A polypeptide sequence composed of 156 amino acids was deduced from the nucleotide sequence. By comparison with the 19 N-terminal amino acids obtained from Edman degradation of the isolated rice 13 kDa prolamin fraction, the signal sequence was determined as consisting of 19 amino acids. The deduced polypeptide is rich in hydrophobic amino acids such as Leu and Val, and also in Gln, but lacks Lys. Hence, the amino acid composition is consistent with that of rice 13 kDa prolamin. By homology with previously reported cereal prolamins, only a single octapeptide sequence, Gln-Gln-Gln-Cys-Cys-Gln-Gln-Leu, which was observed in 15 kDa and 27 kDa zein, B- and gamma-hordein, alpha/beta- and gamma-gliadin, and gamma-secalin was conserved in the rice 10 kDa and 13 kDa prolamin. No repetitive sequences and/or sequences homologous to other cereal prolamins, except the above octapeptide, were observed for the mature 13 kDa prolamin polypeptide. The signal sequence region of the 13 kDa prolamin, however, shows homology of more than 65% in both the nucleotide sequence and the amino acid sequence with rice 10 kDa prolamin and maize zein.
Masumura
T
T
Department of Biochemistry, College of Agriculture, Kyoto Prefectural University, Japan.
Hibino
T
T
Kidzu
K
K
Mitsukawa
N
N
Tanaka
K
K
Fujii
S
S
D000595:Amino Acid Sequence; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics*; D055315:Prolamins; D011506:Proteins / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D010940:Plant Proteins; D055315:Prolamins; D011506:Proteins; D004247:DNA