Bioscience, Biotechnology, and Biochemistry

ISSN: 0916-8451 (Print) 1347-6947 (Online) Journal homepage: http://www.tandfonline.com/loi/tbbb20

Nucleotide Sequence of a cDNA That Encodes a Rice Prolamin Hiroshi Yamagata, Toshiyuki Nomura, Satoshi Arai, Kunisuke Tanaka & Teruo Iwasaki To cite this article: Hiroshi Yamagata, Toshiyuki Nomura, Satoshi Arai, Kunisuke Tanaka & Teruo Iwasaki (1992) Nucleotide Sequence of a cDNA That Encodes a Rice Prolamin, Bioscience, Biotechnology, and Biochemistry, 56:3, 537-537, DOI: 10.1271/bbb.56.537 To link to this article: http://dx.doi.org/10.1271/bbb.56.537

Published online: 12 Jun 2014.

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Date: 05 June 2016, At: 11:44

Biosci. Biotech. Biochem., 56 (3), 537, 1992

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Nucleotide Sequence of a cDNA That Encodes a Rice Prolamin Hiroshi YAMAGATA, Toshiyuki NOMURA, Satoshi ARAI, Kunisuke TANAKA,* and Teruo IWASAKI Laboratory of Biochemistry, Faculty of Agriculture, Kobe University, Nada, Kobe 657, Japan College of Agriculture, Kyoto Prefectural University, Shimogamo, Kyoto 606, Japan Received October 14, 1991

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*Laboratory of Biochemistry,

Rice seed prolamins, alcohol-soluble storage proteins, roughly consist of three polypeptide groups, 10-, 13- and 16-kD ones, which all accumulate in protein body I during endosperm development. 1) Of these prolamin polypeptides, the 13-kD one is the major component. 2 ) The 13-kD prolamin polypeptide and its mRNA show microheterogeneity,3,4) and Southern blot analysis of rice germ DNA suggested that the number of genes for the 13-kD prolamins is large (7-10 copies/haploid genomes) even when hybridization is done under higher stringency conditions. S) Recently, cDNA sequences that encode 13-kD prolamin polypeptides have been reported. S -7) Here we report the nucleotide sequence of a cDNA encoding a previously undescribed 13-kD prolamin polypeptide. A cDNA library constructed in the Agt 11 expression vector from immature rice endosperm poly (A)+ RNA was used for screening for 13-kD prolamin cDNA clones as described previously.S) A cDNA clone (2RM2) was isolated and subcloned into pBluescript SK(+) (Stratagene) at the EcoRI site. The cDNA insert of ARM2 could be split into two fragments of 327 and 274 bp by cleavage with HaeIII (Fig. 1), and these two fragments were also subcloned into the EcoRI and EcoRV sites of pBluescript SK.(+) The complete nucleotide sequences of double-stranded cDNA inserts were analyzed from both directions with the" supercoil sequencing" method. 8 ) The nucleotide sequence (601 bp) and the deduced amino acid sequence (151 residues) are shown in Fig. 2. The signal sequence cleavage site of the precursor polypeptide was suggested to be at Ala 19 _Gln 20 , since the sequence that is identical to the N-terminal amino acid sequence of the isolated l3-kD mature prolamin fractions S) appears from Gln 20 in the deduced amino acid sequence (Fig. 2). These results indicate that ARM2 encodes a major 13-kD prolamin polypeptide and the mature polypeptide consists of 132 amino acids with a molecular mass of 15,059 daltons. The amino acid sequence of the 13-kD prolamin encoded by ARM2 clone was compared with other rice prolamin sequences by the computer programs of IDEAS developed by Kanehisa et al. 9 ) The amino acid sequence is identical at 76%, 85%, 59%, 69%, 83%, and 93% of the alignment positions when compared to the sequences encoded by clones Hae III I

ATTATACAACAAAAATTTAAAAGAACTAGTGTCCTGCAACA

41

ATGAAGATCATTTTCGTCTTTGCTCTCCTTGCTATTGCTGCATGCAGCGCCACTGCGCAG M K I I F V F ALL A I A A C S A T A tiL-

101

1

TTTGATGTTTTAGGTCAAAATATTAGGCAATATCAGGTGCAGTCGCCTCTCCTGCTACAG F D V L G Q N IRQ Y Q v Q S P L L L Q

161

21

CAACAGGTGCTTAGCCCATATAATGAGTTCGTAAGGCAGCAGTATAGCATTGCGGCAAGC Q Q v L Spy N E F V R Q Q Y S I A A S

221

41

ACCTTCTTGCAATCAGCTGCGTTTCAACTGAGAAACAACCAAGTCTTGCAACAGCTCAGG

281

T

CTGGTGGCGCAACAATCTCACTACCAGGACATTAACGTTGTCCAGGCCATAGCGCACCAG L V A Q Q SHY Q D I N V v Q A I A H Q

341

81

CTACACCTCCAGCAGTTTGGCAATCTCTACATTGACCGGAATCTGGCTCAAGCTCAAGCA L H L Q Q F G N L Y I D R N L A Q A Q A

401

101

CTGTTGGCTTTTAACTTGCCATCTACATATGGTATCTACCCTTGGTCCTATAGTGCACCC L L A F N L PST Y G I Y P W S Y SAP

461

121

GATAGCATTACCACCCTTGGCGGTGTCTTGTACTGAATTTTCACAATATTGTAGTTCGGA D SIT T L G G V L Y

521

141

AGTGAAAATATAAGCTCAGGTATCATCGTATGTGACATGTGAAACTTGAGGTGATATAAA

581

TAGAAATAAAATTATCTTTC(An)

601

L

Q

S

A

A

F

Q

L

R

N

N

Q

L

Q

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L

R

The arrow indicates the cleavage site of the signal sequence. Underline below the amino acid sequence indicates that the sequence is identical to the N-terminal amino acid sequence of the mature l3-kd prolamin peptides S ) and underline below the nucleotide sequence indicates the putative polyadenylation sequence.

pProl 7, pProl 14,pProl 17,6) ARM7,5) pX24, and pS23/) respectively.

References 1) 2)

4) 5)

The boxed region encodes an open reading frame.· The hatched region indicates the signal peptide. The direction and extent .of sequence determinations are shown by horizontal arrows.

F

Fig. 2. Nucleotide and Deduced Amino Acid Sequences of the Rice 13-kD Prolamin cDNA Clone ARM2.

3)

Fig. 1. Sequencing Strategy of Rice 13-kDProlamin cDNA Clone ARM2.

v

61

6) 7) 8) 9)

H. Yamagata and K. Tanaka, Plant Cell Physiol., 27, 135-145 (1986). H. Yamagata, T. Sugimoto, K. Tanaka, and Z. Kasai, Plant Physiol., 70, 1094-1100 (1982). T. Hibino, K. Kidzu, T. Masumura, K. Ohtsuka, K. Tanaka, M. Kawabata, and S. Fujii, Agric. Bioi. Chern., 53, 5IJ--518 (1989). H. Yamagata, K. Tamura, K. Tanaka, and Z. Kasai, Plant Cell Physiol., 27, 1419-1422 (1986). T. Masumura, T. Hibino, K. Kidzu, N. Mitsukawa, K. Tanaka, and S. Fujii, Mol. Gen. Gene!;, 221, 1-7 (1990). W. T. Kim and T. W. Okita, Plant Physiol., 88, 649-655 (1988). L. F. Shyur and C. S. Chen, Nucl. Acids Res., 18, 6683 (1990). E. Y. Chen and P.R. Seeburg, DNA, 4,165-170 (1985). M.I

Nucleotide sequence of a cDNA that encodes a rice prolamin.

Bioscience, Biotechnology, and Biochemistry ISSN: 0916-8451 (Print) 1347-6947 (Online) Journal homepage: http://www.tandfonline.com/loi/tbbb20 Nucle...
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