Skip to main content
Plant Physiology logoLink to Plant Physiology
. 1993 Mar;101(3):809–817. doi: 10.1104/pp.101.3.809

Isolation and characterization of a cDNA clone for plant nuclear antigen 21D7 associated with cell division.

M W Smith 1, M Ito 1, T Yamada 1, T Suzuki 1, A Komamine 1
PMCID: PMC158694  PMID: 8310059

Abstract

A cDNA clone was isolated from a carrot (Daucus carota L.) cDNA expression library using monoclonal antibody 21D7, which recognizes a nuclear antigen associated with cell division in plant cells. To show that the isolated cDNA encodes the 21D7 antigen, a polyclonal antiserum was raised against a recombinant fusion protein specified by the cDNA. Both the polyclonal antiserum and the monoclonal antibody 21D7 recognized the same plant protein on immunoblots, in immunoprecipitation experiments, and in peptide mapping. Analysis of the cDNA revealed that the deduced amino acid sequence has 45% identity to the predicted sequence of the mouse transplantation antigen P91A from mutant tumor cells that is responsible for the immune rejection of the corresponding cell clone in a syngeneic mouse. The expression of the plant cDNA at the mRNA level was highly correlated with cell proliferation. In suspension cultures of Catharanthus roseus (L.) G Don. cells, the highest level of expression was observed during the midlogarithmic phase of growth. When auxin was added to stimulate cell division of auxin-starved cells arrested in the G1 phase, transcription was immediately enhanced, and the level of expression remained high throughout the G1 and S phases and dropped dramatically at the end of DNA replication.

Full Text

The Full Text of this article is available as a PDF (2.5 MB).

Selected References

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

  1. Boon T. Antigenic tumor cell variants obtained with mutagens. Adv Cancer Res. 1983;39:121–151. doi: 10.1016/s0065-230x(08)61034-9. [DOI] [PubMed] [Google Scholar]
  2. Boon T., Van Pel A. T cell-recognized antigenic peptides derived from the cellular genome are not protein degradation products but can be generated directly by transcription and translation of short subgenic regions. A hypothesis. Immunogenetics. 1989;29(2):75–79. doi: 10.1007/BF00395854. [DOI] [PubMed] [Google Scholar]
  3. Carrington J. C., Freed D. D., Leinicke A. J. Bipartite signal sequence mediates nuclear translocation of the plant potyviral NIa protein. Plant Cell. 1991 Sep;3(9):953–962. doi: 10.1105/tpc.3.9.953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  5. Dingwall C., Laskey R. A. Nuclear targeting sequences--a consensus? Trends Biochem Sci. 1991 Dec;16(12):478–481. doi: 10.1016/0968-0004(91)90184-w. [DOI] [PubMed] [Google Scholar]
  6. Garcia-Bustos J., Heitman J., Hall M. N. Nuclear protein localization. Biochim Biophys Acta. 1991 Mar 7;1071(1):83–101. doi: 10.1016/0304-4157(91)90013-m. [DOI] [PubMed] [Google Scholar]
  7. Hirt H., Páy A., Györgyey J., Bakó L., Németh K., Bögre L., Schweyen R. J., Heberle-Bors E., Dudits D. Complementation of a yeast cell cycle mutant by an alfalfa cDNA encoding a protein kinase homologous to p34cdc2. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1636–1640. doi: 10.1073/pnas.88.5.1636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ito M., Kodama H., Komamine A. Identification of a novel S-phase-specific gene during the cell cycle in synchronous cultures of Catharanthus roseus cells. Plant J. 1991 Sep;1(2):141–148. doi: 10.1111/j.1365-313x.1991.00141.x. [DOI] [PubMed] [Google Scholar]
  9. Jacobs T. Control of the cell cycle. Dev Biol. 1992 Sep;153(1):1–15. doi: 10.1016/0012-1606(92)90087-w. [DOI] [PubMed] [Google Scholar]
  10. Kodama H., Ito M., Ohnishi N., Suzuka I., Komamine A. Molecular cloning of the gene for plant proliferating-cell nuclear antigen and expression of this gene during the cell cycle in synchronized cultures of Catharanthus roseus cells. Eur J Biochem. 1991 Apr 23;197(2):495–503. doi: 10.1111/j.1432-1033.1991.tb15937.x. [DOI] [PubMed] [Google Scholar]
  11. Luna E. J., Kidd G. H., Branton D. Identification by peptide analysis of the spectrin-binding protein in human erythrocytes. J Biol Chem. 1979 Apr 10;254(7):2526–2532. [PubMed] [Google Scholar]
  12. McKinney J. D., Heintz N. Transcriptional regulation in the eukaryotic cell cycle. Trends Biochem Sci. 1991 Nov;16(11):430–435. doi: 10.1016/0968-0004(91)90170-z. [DOI] [PubMed] [Google Scholar]
  13. Moreland R. B., Langevin G. L., Singer R. H., Garcea R. L., Hereford L. M. Amino acid sequences that determine the nuclear localization of yeast histone 2B. Mol Cell Biol. 1987 Nov;7(11):4048–4057. doi: 10.1128/mcb.7.11.4048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Silver P. A. How proteins enter the nucleus. Cell. 1991 Feb 8;64(3):489–497. doi: 10.1016/0092-8674(91)90233-o. [DOI] [PubMed] [Google Scholar]
  15. Takahashi Y., Kusaba M., Hiraoka Y., Nagata T. Characterization of the auxin-regulated par gene from tobacco mesophyll protoplasts. Plant J. 1991 Nov;1(3):327–332. doi: 10.1046/j.1365-313x.1991.t01-2-00999.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES