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. 1985 May;4(5):1103–1110. doi: 10.1002/j.1460-2075.1985.tb03746.x

Each zein gene class can produce polypeptides of different sizes

A Viotti 1, G Cairo 1, A Vitale 1, E Sala 1
PMCID: PMC554310  PMID: 16453609

Abstract

The family of zein proteins in maize consists of several prototypes whose amino sequence is characterized by a repetitive block structure. We have now isolated and characterized a new series of zein cDNA clones from a cDNA library made from protein-body polysomal mRNA. The nucleotide sequences and the hybrid-selected translation results indicate that anomalous zein genes with termination codons within the coding region are transcribed, translated and possibly accumulated into zein protein bodies. The sequence analysis of these new clones and their comparison with already characterized zein sequences, all from the same maize line, indicate a large variability in the amino acid sequence of the blocks both among the prototypes and within members of each prototype. Northern blot analysis of total RNA from endosperms at different maturation stages shows a differential expression of each prototype sequence with the production, within each family, of zein transcripts coding for discrete size classes of polypeptides. The results allow a better definition of the zein system, clarify the relationship between sequence types and polypeptide size classes, and suggest the possibility of inserting lysine codons to raise the nutritional value of the seed storage protein.

Keywords: zein prototypes, in-frame stop codons, truncated polypeptides, timing of expression

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Selected References

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  1. Argos P., Pedersen K., Marks M. D., Larkins B. A. A structural model for maize zein proteins. J Biol Chem. 1982 Sep 10;257(17):9984–9990. [PubMed] [Google Scholar]
  2. Burr B., Burr F. A. Zein synthesis in maize endosperm by polyribosomes attached to protein bodies. Proc Natl Acad Sci U S A. 1976 Feb;73(2):515–519. doi: 10.1073/pnas.73.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  4. Geraghty D. E., Messing J., Rubenstein I. Sequence analysis and comparison of cDNAs of the zein multigene family . EMBO J. 1982;1(11):1329–1335. doi: 10.1002/j.1460-2075.1982.tb01318.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
  6. Hagen G., Rubenstein I. Complex organization of zein genes in maize. Gene. 1981 Apr;13(3):239–249. doi: 10.1016/0378-1119(81)90029-9. [DOI] [PubMed] [Google Scholar]
  7. Hague D. R. Studies of storage proteins of higher plants: I. Concanavalin a from three species of the genus canavalia. Plant Physiol. 1975 Apr;55(4):636–642. doi: 10.1104/pp.55.4.636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Heidecker G., Messing J. Sequence analysis of zein cDNAs obtained by an efficient mRNA cloning method. Nucleic Acids Res. 1983 Jul 25;11(14):4891–4906. doi: 10.1093/nar/11.14.4891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Langridge P., Feix G. A zein gene of maize is transcribed from two widely separated promoter regions. Cell. 1983 Oct;34(3):1015–1022. doi: 10.1016/0092-8674(83)90559-7. [DOI] [PubMed] [Google Scholar]
  11. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  12. Marks M. D., Larkins B. A. Analysis of sequence microheterogeneity among zein messenger RNAs. J Biol Chem. 1982 Sep 10;257(17):9976–9983. [PubMed] [Google Scholar]
  13. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  14. Pedersen K., Devereux J., Wilson D. R., Sheldon E., Larkins B. A. Cloning and sequence analysis reveal structural variation among related zein genes in maize. Cell. 1982 Jul;29(3):1015–1026. doi: 10.1016/0092-8674(82)90465-2. [DOI] [PubMed] [Google Scholar]
  15. Salamini F. Controlling elements at the opaque-2 locus of maize: their involvement in the origin of spontaneous mutation. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 2):467–476. doi: 10.1101/sqb.1981.045.01.062. [DOI] [PubMed] [Google Scholar]
  16. Spena A., Viotti A., Pirrotta V. A homologous repetitive block structure underlies the heterogeneity of heavy and light chain zein genes. EMBO J. 1982;1(12):1589–1594. doi: 10.1002/j.1460-2075.1982.tb01360.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Spena A., Viotti A., Pirrotta V. Two adjacent genomic zein sequences: structure, organization and tissue-specific restriction pattern. J Mol Biol. 1983 Oct 5;169(4):799–811. doi: 10.1016/s0022-2836(83)80137-5. [DOI] [PubMed] [Google Scholar]
  18. Steele J. C., Jr, Nielsen T. B. Evaluation of cross-linked polypeptides in SDS gel electrophoresis. Anal Biochem. 1978 Jan;84(1):218–224. doi: 10.1016/0003-2697(78)90502-x. [DOI] [PubMed] [Google Scholar]
  19. Szalay A. A., Grohmann K., Sinsheimer R. L. Separation of the complementary strands of DNA fragments on polyacrylamide gels. Nucleic Acids Res. 1977;4(5):1569–1578. doi: 10.1093/nar/4.5.1569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Talbot D. N., Yphantis D. A. Fluorescent monitoring of SDS gel electrophoresis. Anal Biochem. 1971 Nov;44(1):246–253. doi: 10.1016/0003-2697(71)90367-8. [DOI] [PubMed] [Google Scholar]
  21. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  22. Viotti A., Abildsten D., Pogna N., Sala E., Pirrotta V. Multiplicity and diversity of cloned zein cDNA sequences and their chromosomal localisation. EMBO J. 1982;1(1):53–58. doi: 10.1002/j.1460-2075.1982.tb01123.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Viotti A., Balducci C., Weil J. H. Adaptation of the tRNA population of maize endosperm for zein synthesis. Biochim Biophys Acta. 1978 Jan 26;517(1):125–132. doi: 10.1016/0005-2787(78)90040-0. [DOI] [PubMed] [Google Scholar]
  24. Viotti A., Sala E., Marotta R., Alberi P., Balducci C., Soave C. Genes and mRNAs coding for zein polypeptides in Zea mays. Eur J Biochem. 1979 Dec;102(1):211–222. doi: 10.1111/j.1432-1033.1979.tb06282.x. [DOI] [PubMed] [Google Scholar]
  25. Wang J. L., Cunningham B. A., Edelman G. M. Unusual fragments in the subunit structure of concanavalin A. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1130–1134. doi: 10.1073/pnas.68.6.1130. [DOI] [PMC free article] [PubMed] [Google Scholar]

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