Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1989 Sep;91(1):124–129. doi: 10.1104/pp.91.1.124

Cloning and Characterization of Root-Specific Barley Lectin 1

David R Lerner 1, Natasha V Raikhel 1
PMCID: PMC1061962  PMID: 16666982

Abstract

Cereal lectins are a class of biochemically and antigenically related proteins localized in a tissue-specific manner in embryos and adult plants. To study the specificity of lectin expression, a barley (Hordeum vulgare L.) embryo cDNA library was constructed and a clone (BLc3) for barley lectin was isolated. BLc3 is 972 nucleotides long and includes an open reading frame of 212 amino acids. The deduced amino acid sequence contains a putative signal peptide of 26 amino acid residues followed by a 186 amino acid polypeptide. This polypeptide has 95% sequence identity to the antigenically indistinguishable wheat germ agglutinin isolectin-B (WGA-B) suggesting that BLc3 encodes barley lectin. Further evidence that BLc3 encodes barley lectin was obtained by immunoprecipitation of the in vitro translation products of BLc3 RNA transcripts and barley embryo poly(A+) RNA. In situ hybridizations with BLc3 showed that barley lectin gene expression is confined to the outermost cell layers of both embryonic and adult root tips. On Northern blots, BLc3 hybridizes to a 1.0 kilobyte mRNA in poly(A+) RNA from both embryos and root tips. We suggest, on the basis of immunoblot experiments, that barley lectin is synthesized as a glycosylated precursor and processed by removal of a portion of the carboxyl terminus including the single N-linked glycosylation site.

Full text

PDF
124

Images in this article

126Figure 2
on p.126
126Figure 3
on p.126
127Figure 4
on p.127
128Figure 5
on p.128

Selected References

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

  1. Dale R. M., Arrow A. A rapid single-stranded cloning, sequencing, insertion, and deletion strategy. Methods Enzymol. 1987;155:204–214. doi: 10.1016/0076-6879(87)55017-0. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Fett W. F., Dunn M. F. Exopolysaccharides Produced by Phytopathogenic Pseudomonas syringae Pathovars in Infected Leaves of Susceptible Hosts. Plant Physiol. 1989 Jan;89(1):5–9. doi: 10.1104/pp.89.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Finkelstein R. R., Crouch M. L. Rapeseed embryo development in culture on high osmoticum is similar to that in seeds. Plant Physiol. 1986 Jul;81(3):907–912. doi: 10.1104/pp.81.3.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  6. Mishkind M. L., Palevitz B. A., Raikhel N. V., Keegstra K. Localization of wheat germ agglutinin--like lectins in various species of the gramineae. Science. 1983 Jun 17;220(4603):1290–1292. doi: 10.1126/science.220.4603.1290. [DOI] [PubMed] [Google Scholar]
  7. Mishkind M., Raikhel N. V., Palevitz B. A., Keegstra K. Immunocytochemical localization of wheat germ agglutinin in wheat. J Cell Biol. 1982 Mar;92(3):753–764. doi: 10.1083/jcb.92.3.753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Mizusawa S., Nishimura S., Seela F. Improvement of the dideoxy chain termination method of DNA sequencing by use of deoxy-7-deazaguanosine triphosphate in place of dGTP. Nucleic Acids Res. 1986 Feb 11;14(3):1319–1324. doi: 10.1093/nar/14.3.1319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Peumans W. J., Stinissen H. M., Carlier A. R. Isolation and partial characterization of wheat-germ-agglutinin-like lectins from rye (Secale cereale) and barley (Hordeum vulgare) embryos. Biochem J. 1982 Apr 1;203(1):239–243. doi: 10.1042/bj2030239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Raikhel N. V., Wilkins T. A. Isolation and characterization of a cDNA clone encoding wheat germ agglutinin. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6745–6749. doi: 10.1073/pnas.84.19.6745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Shinshi H., Wenzler H., Neuhaus J. M., Felix G., Hofsteenge J., Meins F. Evidence for N- and C-terminal processing of a plant defense-related enzyme: Primary structure of tobacco prepro-beta-1,3-glucanase. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5541–5545. doi: 10.1073/pnas.85.15.5541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Silflow C. D., Hammett J. R., Key J. L. Sequence complexity of polyadenylated ribonucleic acid from soybean suspension culture cells. Biochemistry. 1979 Jun 26;18(13):2725–2731. doi: 10.1021/bi00580a006. [DOI] [PubMed] [Google Scholar]
  14. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Triplett B. A., Quatrano R. S. Timing, localization, and control of wheat germ agglutinin synthesis in developing wheat embryos. Dev Biol. 1982 Jun;91(2):491–496. doi: 10.1016/0012-1606(82)90057-4. [DOI] [PubMed] [Google Scholar]
  16. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
  17. Wilkins T. A., Raikhel N. V. Expression of rice lectin is governed by two temporally and spatially regulated mRNAs in developing embryos. Plant Cell. 1989 May;1(5):541–549. doi: 10.1105/tpc.1.5.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES