Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1988 Feb 15;250(1):15–24. doi: 10.1042/bj2500015

Two genes encoding 'minor' legumin polypeptides in pea (Pisum sativum L.). Characterization and complete sequence of the LegJ gene.

J A Gatehouse 1, D Bown 1, J Gilroy 1, M Levasseur 1, J Castleton 1, T H Ellis 1
PMCID: PMC1148808  PMID: 3355508

Abstract

A genomic clone from pea (Pisum sativum L.) contains all of one gene encoding a 'minor' (B-type) legumin polypeptide, and most of a second very similar gene. The two genes, designated LegJ and LegK, are arranged in tandem, separated by approx. 6 kb. A complete sequence of gene LegJ and its flanking sequences is given, with as much of the sequence of gene LegK as is present on the genomic clone. Hybridization of 3' flanking sequence probes to seed mRNA, and sequence comparisons with cDNA species, suggested that gene LegJ, and probably gene LegK, was expressed. The partial amino acid sequences of 'minor' legumin alpha- and beta-polypeptides were used to confirm the identity of these genes. The transciption start in gene LegJ was mapped. The 5' flanking sequence of gene LegJ contains a sequence conserved in legumin genes from pea and other species, which is likely to have functional significance in control of gene expression. Sequence comparisons with legumin genes and cDNA species from Vicia faba and soya bean show that separation of legumin genes into A- and B-type subfamilies occurred before separation of the Viciae and Glycinae tribes.

Full text

PDF
19

Images in this article

20Fig. 3.
on p.20
21Fig. 4.
on p.21

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bown D., Levasseur M., Croy R. R., Boulter D., Gatehouse J. A. Sequence of a pseudogene in the legumin gene family of pea (Pisum sativum L.). Nucleic Acids Res. 1985 Jun 25;13(12):4527–4538. doi: 10.1093/nar/13.12.4527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Breathnach R., Benoist C., O'Hare K., Gannon F., Chambon P. Ovalbumin gene: evidence for a leader sequence in mRNA and DNA sequences at the exon-intron boundaries. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4853–4857. doi: 10.1073/pnas.75.10.4853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bäumlein H., Wobus U., Pustell J., Kafatos F. C. The legumin gene family: structure of a B type gene of Vicia faba and a possible legumin gene specific regulatory element. Nucleic Acids Res. 1986 Mar 25;14(6):2707–2720. doi: 10.1093/nar/14.6.2707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  6. Domoney C., Casey R. Measurement of gene number for seed storage proteins in Pisum. Nucleic Acids Res. 1985 Feb 11;13(3):687–699. doi: 10.1093/nar/13.3.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Domoney C., Casey R. Storage protein precursor polypeptides in cotyledons of Pisum sativum L. Identification of, and isolation of a cDNA clone for, an 80000-Mr legumin-related polypeptide. Eur J Biochem. 1984 Mar 1;139(2):321–327. doi: 10.1111/j.1432-1033.1984.tb08010.x. [DOI] [PubMed] [Google Scholar]
  8. Dynan W. S., Tjian R. Control of eukaryotic messenger RNA synthesis by sequence-specific DNA-binding proteins. 1985 Aug 29-Sep 4Nature. 316(6031):774–778. doi: 10.1038/316774a0. [DOI] [PubMed] [Google Scholar]
  9. Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
  10. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  11. Fukazawa C., Momma T., Hirano H., Harada K., Udaka K. Glycinin A3B4 mRNA. Cloning and sequencing of double-stranded cDNA complementary to a soybean storage protein. J Biol Chem. 1985 May 25;260(10):6234–6239. [PubMed] [Google Scholar]
  12. Gatehouse J. A., Croy R. D., Boulter D. Isoelectric-focusing properties and carbohydrate content of pea (Pisum sativum) legumin. Biochem J. 1980 Feb 1;185(2):497–503. doi: 10.1042/bj1850497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gatehouse J. A., Evans I. M., Bown D., Croy R. R., Boulter D. Control of storage-protein synthesis during seed development in pea (Pisum sativum L.). Biochem J. 1982 Oct 15;208(1):119–127. doi: 10.1042/bj2080119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gatehouse J. A., Lycett G. W., Croy R. R., Boulter D. The post-translational proteolysis of the subunits of vicilin from pea (Pisum sativum L.). Biochem J. 1982 Dec 1;207(3):629–632. doi: 10.1042/bj2070629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gates M. A. Simpler DNA sequence representations. Nature. 1985 Jul 18;316(6025):219–219. doi: 10.1038/316219a0. [DOI] [PubMed] [Google Scholar]
  16. Hoffman L. M., Donaldson D. D. Characterization of two Phaseolus vulgaris phytohemagglutinin genes closely linked on the chromosome. EMBO J. 1985 Apr;4(4):883–889. doi: 10.1002/j.1460-2075.1985.tb03714.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kühn S., Fritz H. J., Starlinger P. Close vicinity of IS1 integration sites in the leader sequence of the gal operon of E. coli. Mol Gen Genet. 1979 Jan 2;167(3):235–241. doi: 10.1007/BF00267414. [DOI] [PubMed] [Google Scholar]
  18. Lycett G. W., Croy R. R., Shirsat A. H., Boulter D. The complete nucleotide sequence of a legumin gene from pea (Pisum sativum L.). Nucleic Acids Res. 1984 Jun 11;12(11):4493–4506. doi: 10.1093/nar/12.11.4493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lütcke H. A., Chow K. C., Mickel F. S., Moss K. A., Kern H. F., Scheele G. A. Selection of AUG initiation codons differs in plants and animals. EMBO J. 1987 Jan;6(1):43–48. doi: 10.1002/j.1460-2075.1987.tb04716.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marco Y. A., Thanh V. H., Tumer N. E., Scallon B. J., Nielsen N. C. Cloning and structural analysis of DNA encoding an A2B1a subunit of glycinin. J Biol Chem. 1984 Nov 10;259(21):13436–13441. [PubMed] [Google Scholar]
  21. 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]
  22. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  24. Momma T., Negoro T., Hirano H., Matsumoto A., Udaka K., Fukazawa C. Glycinin A5A4B3 mRNA: cDNA cloning and nucleotide sequencing of a splitting storage protein subunit of soybean. Eur J Biochem. 1985 Jun 18;149(3):491–496. doi: 10.1111/j.1432-1033.1985.tb08951.x. [DOI] [PubMed] [Google Scholar]
  25. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  26. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Staden R. An interactive graphics program for comparing and aligning nucleic acid and amino acid sequences. Nucleic Acids Res. 1982 May 11;10(9):2951–2961. doi: 10.1093/nar/10.9.2951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wright D. J., Boulter D. Purification and subunit structure of legumin of Vicia faba L. (broad bean). Biochem J. 1974 Aug;141(2):413–418. doi: 10.1042/bj1410413. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES