The sequence of the zein regulatory gene opaque-2 (O2) of Zea Mays.
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Biomedical subjects
Publications and source records attributed to M Motto.
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The structure of the zein regulatory gene Opaque 2 of Zea mays has been determined by sequence analysis of genomic and cDNA clones. The size of O2 mRNA is 1751 bp [poly(A) tail not included] containing a major open reading frame (ORF) of 1380 bp preceded by three short ORFs of 3, 21 and 20 amino acid residues. The main ORF comprises 1362 bp and is composed of six exons ranging in size from 465 to 61 bp and five introns of 678 bp to 83 bp. A putative protein 454 amino acids long was derived by the theoretical translation of the genomic sequences corresponding to exons. The opaque 2 protein contains a domain similar to the leucine zipper motif identified in DNA binding proteins of animal protooncogenes such as fos, jun and myc, and in the transcriptional activators GCN4 and C/EBP. The region of 30 amino acid residues next to the leucine repeats towards the N terminus is rich in basic amino acids and is also homologous to a domain present in fos, jun and GCN4. Moreover, in the carboxy terminal region an amino acid motif closely resembling a metal binding domain is present.
The cDNA coding for the b-32 protein, an albumin expressed in maize endosperm cells under the control of the O2 and O6 loci, has been cloned and the complete amino acid sequence of the protein derived. A lambda gt11 cDNA library from mRNA of immature maize endosperm was screened for the expression of the b-32 protein using antibodies against the purified protein. One of the positive clones obtained was used to isolate a full-length cDNA clone. By Northern analysis, the size of the b-32 mRNA was estimated to be 1.2 kb. Hybrid-selected translation assays show that the message codes for a protein with an apparent molecular weight of 30-35 kDa. The nucleotide sequence shows that several internal repeats are present. The protein has a length of 303 amino acid residues (mol. wt. 32430 dalton) and its sequence shows the following features: no signal peptide is observable; it contains seven tryptophan residues, an amino acid absent in maize storage proteins; polar and hydrophobic residues are spread along the sequence; several pairs of basic residues are present in the N-terminal region; the secondary structure allows the prediction of two structural domains for the b-32 protein that would fold up giving rise to a globular shape. The cloning of this gene may help in understanding the role of the O2 and O6 loci in regulating the deposition of zein, the major storage protein of maize endosperm.
Objective of research was (a) to evaluate the influence of pollination-prevention on various metabolic parameters of the two maize inbreds B73 and B14A and their F1, and (b) to gain information on the inheritance of leaf senescence, in response to pollination-prevention. The results show that the visual pattern of leaf senescence, in response to prevention of ear pollination, contrasts markedly between the two inbred lines. Relative to control plants, prevention of ear pollination, causes a premature senescence in B73 and B73 x B14A plants, while leaves of unpollinated B14A remain green and similar in appearance to pollinated controls. Furthermore, prevention of ear pollination induces a sizable reduction of dry matter accumulation of all above-ground material and changes in various metabolic parameters. An accumulation of sucrose in the leaves of unpollinated B73 and B73 x B14A plants is correlated with the development of premature senescence. Finally, the genetic analysis supports suggestions that a single dominant gene is responsible for the differences observed, in the visual pattern of leaf senescence, in response to prevention of ear pollination.
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