Abstract
A cDNA clone, pCD7, was shown to hybrid-select from developing seeds of Pisum sativum L. an mRNA that translated into a polypeptide of apparent Mr 90 000. The translation product was observed only in the earliest stages of embryogenesis and was detected at a developmental stage when virtually all the cotyledon cells are mitotic. Sequence analysis of pCD7 showed it to correspond to a member of the 70 000-Mr heat-shock protein (HSP70) gene family. Transcripts corresponding to pCD7 were detected in different P. sativum organs, with roots apparently showing lower levels of pCD7-homologous RNA than other organs. Hybridizations to P. sativum DNA identified polymorphisms in the genomic DNA corresponding to pCD7 and the segregation of these in selected crosses indicated the existence of at least two genetic loci, one of which mapped to an existing linkage group.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- cDNA:
-
complementary DNA
- DAF:
-
days after flowering
- HSC:
-
heat-shock cognate
- HSP:
-
heat-shock protein
- SDS:
-
sodium dodecyl sulphate
- SSC:
-
0.15 M sodium chloride, 0.015 M sodium citrate, pH 7.0
References
Ambrose, M.J., Wang, T.L., Cook, S.K., Hedley, C.L. (1987) An analysis of seed development in Pisum sativum cell populations in vivo and in vitro. J. Exp. Bot. 38, 1909–1920
Ajir-Shapira, D., Leustek, T., Dalie, B., Weissbach, H., Brot, N. (1990) Hsp70 proteins, similar to Escherichia coli DnaK, in chloroplasts and mitochondria of Euglena gracilis. Proc. Natl. Acad. Sci. USA 87, 1749–1752
Bardwell, J.C.A., Craig, E.A. (1984) Major heat shock gene of Drosophila and the Escherichia coli heat-inducible dnaK gene are homologous. Proc. Natl. Acad. Sci. USA 81, 848–852
Biggin, M.D., Gibson, T.J., Hong, G.F. (1983) Buffer gradient gels and −35S label as an aid to rapid DNA sequence determination. Proc. Natl. Acad. Sci. USA 80, 3963–3965
Devereux, J., Haeberli, P., Smithies, O. (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 12, 387–395
Domoney, C., Casey, R. (1983) Cloning and characterization of complementary DNA for convicilin, a major seed storage protein in Pisum sativum L.. Planta 159, 446–453
Domoney, C., Casey, R. (1985) Measurement of gene number for seed storage proteins in Pisum. Nucl. Acids Res. 13, 687–699
Domoney, C., Casey, R. (1987) Changes in legumin messenger RNAs throughout seed development in Pisum sativum L.. Planta 170, 562–566
Domoney, C., Ellis, T.H.N., Davies, D.R. (1986) Organization and mapping of legumin genes in Pisum. Mol. Gen. Genet. 202, 280–285
Domoney, C., Firmnin, J.L., Sidebottom C., Ealing, P.M., Slabas, A., Casey, R. (1990) Lipoxygenase heterogeneity in Pisum sativum. Planta 181, 35–43
Duck, N., McCormick, S., Winter, J. (1989) Heat shock protein hsp70 cognate gene expression in vegetative and reproductive organs of Lycopersicon esculentum. Proc. Natl. Acad. Sci. USA 86, 3674–3678
Ellis, R.J. (1990) The molecular chaperone concept. Semin. Cell Biol. 1, 1–9
Ellis, T.H.N., Davies, D.R., Castleton, J.A., Bedford, I.D. (1984) The organization and genetics of rDNA length variants in peas. Chromosoma (Berl.) 91, 74–81
Ellis, T.H.N., Domoney, C., Castleton, J., Cleary, W., Davies, D.R. (1986) Vicilin genes in Pisum. Mol. Gen. Genet. 205, 164–169
Flaherty, K.M., DeLuca-Flaherty, C., McKay, D.B. (1990) Three-dimensional structure of the ATPase fragment of a 70K heatshock cognate protein. Nature 346, 623–628
Hardesty, B., Kramer, G. (1989) The 90 000 dalton heat shock protein, a lot of smoke but no function as yet. Biochem. Cell. Biol. 67, 749–750
Hemmingsen, S.M., Woolford, C., van der Vies, S.M., Tilly, K., Dennis, D.T., Georgopoulos, C.P., Hendrix R.W., Ellis, R.J. (1988) Homologous plant and bacterial proteins chaperone oligomeric protein assembly. Nature 333, 330–334
Hunt, C., Morimoto, R.I. (1985) Conserved features of eukaryotic hsp70 genes revealed by comparison with the nucleotide sequence of human hsp70. Proc. Natl. Acad. Sci. USA 82, 6455–6459
Ingolia, T.D., Craig, E.A., McCarthy, B.J. (1980) Sequence of three copies of the gene for the major Drosophila heat shock induced protein and their flanking regions. Cell 21, 669–679
Innis, M.A., Myambo, K.B., Gelfand, D.H., Brow, M.A.D. (1988) DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA. Proc. Natl. Acad. Sci. USA 85, 9436–9440
Lindquist, S. (1986) The heat-shock response. Annu. Rev. Biochem. 55, 1151–1191
Lindquist, S., Craig E.A. (1988) The heat shock proteins. Annu. Rev. Genet. 22, 631–677
Marshall, J.S., DeRocher, A.E., Keegstra, K., Vierling, E. (1990) Identification of heat shock protein hsp70 homologues in chloroplasts. Proc. Natl. Acad. Sci. USA 87, 374–378
Nagao, T., Kimpel, J.A., Vierling, E., Key, J.L. (1986) The heat shock response: a comparative analysis. Oxf. Surv. Plant. Mol. Cell Biol. 3, 384–438
Parry-Smith, D.J., Attwood, T.K. (1991) A novel approach to multiple sequence alignment. Computer Applications in Bio Sciences, vol. 7, pt. 2, in press
Pelham, H.R.B. (1986) Speculations on the functions of the major heat shock and glucose-regulated proteins. Cell 46, 959–961
Rochester, D.E., Winer J.A., Shah, D.M. (1986) The structure and expression of maize genes encoding the major heat shock protein, hsp70. EMBO J. 5, 451–458
Sanger, F., Nicklen, S., Coulson, A.R. (1977) DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74, 5463–5467
Sheppard, M., Kemp, D.J., Anders, R.F., Lew, A.M. (1989) High level sequence homology between a Plasmodium chabaudi heat shock protein gene and its Plasmodium falciparum equivalent. Mol. Biochem. Parasitol. 33, 101–103
Staden, R. (1982a) An interactive graphics program for comparing and aligning nucleic acid and amino acid sequences. Nucleic Acids Res. 10, 2951–2961
Staden, R. (1982b) Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing. Nucleic Acids Res. 10, 4731–4751
Tabor, S., Richardson, C.C. (1987) DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc. Natl. Acad. Sci. USA 84, 4767–4771
Wu, C.H., Caspar, T., Browse, J., Lindquist, S., Somerville, C. (1988) Characterization of an HSP70 cognate gene family in Arabidopsis. Plant Physiol. 88, 731–740
Author information
Authors and Affiliations
Additional information
This work was supported by the Agricultural and Food Research council via a grant-in-aid to the John Innes Institute.
Rights and permissions
About this article
Cite this article
Domoney, C., Ellis, N., Turner, L. et al. A developmentally regulated early-embryogenesis protein in pea (Pisum sativum L.) is related to the heat-shock protein (HSP70) gene family. Planta 184, 350–355 (1991). https://doi.org/10.1007/BF00195336
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00195336