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Production of multiple plant hormones from a single polyprotein precursor

Nature volume 411, pages 817–820 (2001)Cite this article

Abstract

Some animal and yeast hormone genes produce prohormone polypeptides that are proteolytically processed to produce multiple copies of hormones with the same or different functions1. In plants, four polypeptides have been identified that can be classed as hormones2,3,4,5 (intercellular chemical messengers6) but none are known to be produced as multiple copies from a single precursor. Here we describe a polyprotein hormone precursor, present in tobacco plants, that gives rise to two polypeptide hormones, as often found in animals and yeast. The tobacco polypeptides activate the synthesis of defensive proteinase-inhibitor proteins in a manner similar to that of systemin, an 18-amino-acid polypeptide found in tomato plants2. The two tobacco polypeptides are derived from each end of a 165-amino-acid precursor that bears no homology to tomato prosystemin. The data show that structurally diverse polypeptide hormones in different plant species can serve similar signalling roles, a condition not found in animals or yeast.

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Figure 1: Isolation of biologically active tobacco polypeptides from tobacco leaf extracts.
Figure 2: The biological activities of polypeptides I and II.
Figure 3: The nucleotide and amino-acid sequence of the Tob Sys I and II polyprotein precursor.
Figure 4: Activation of a MAP kinase with Mr 48K in tobacco suspension-cultured cells by Tob Sys I and II.

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Accession codes

Accessions

GenBank/EMBL/DDBJ

Data deposits

The GenBank accession number for tobacco systemin precursor pro-TobSys-A is AY033148, and for pro-TobSys-B is AY033149.

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Acknowledgements

This research was supported by the College of Agriculture and Home Economics and by the National Science Foundation. We thank S. Vogtman for growing plants; G. Munske for amino-acid sequence analyses; and W. Siems for MALDI mass spectroscopic analyses.

Author information

Author notes

  1. Johannes Stratmann

    Present address: Department of Biological Sciences, University of South Carolina, Columbia, South Carolina, 29208, USA

  2. Clarence A. Ryan: Correspondence and requests for materials should be addressed to C.A.R.

Authors and Affiliations

  1. Institute of Biological Chemistry, Washington State University, Pullman, 99164-6340, Washington, USA

    Gregory Pearce, Daniel S. Moura, Johannes Stratmann & Clarence A. Ryan

Authors
  1. Gregory Pearce
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Corresponding author

Correspondence to Clarence A. Ryan.

Supplementary information

Supplement 1.

(DOC 23 KB) Protocol for the purification of tobacco systemins

Supplement 2.

(JPG 17 KB)

Alkalinization of the medium of suspension cultured tobacco cells in response to increasing concentrations of the ‘crude polypeptide extract’ from leaves as described in the Methods

Supplement 3.

(PPT 378 KB)

The changes in molecular masses of Tob Sys I and II during the hydrolysis of 100 pmoles of each in 100 mL 1% trifluoroacetic acid at 80 °C. At the intervals shown, 10 μL aliquots were removed for MALDI-MS analysis. A ladder of fragments duffering by 132 mass units (indicative of pentoses) were produced as the carbohydrate moieties were removed from the polypeptides. a. 0 min. b. 15 min. c. 30 min d. 60 min. e. 180 min.

Supplement 4.

(JPG 38 KB)

Alkalinization assays of synthetic Tob Sys I and II backbone polypeptides using tobacco suspension cultured cells. The change in pH of the culture medium was measured 15 min after addition of the polypeptides. Half maximal activity of each polypeptide is about 2 μM, as compared to 200 pM of the native polypeptides (cf. Figure 2a).

Supplement 5.

(JPG 67 KB)

Alignment of the two deduced tobacco systemin precursor proteins (pro-TobSys-A and pro-TobSys-B). The alignment was made using the Genetics Computer Guoup (GCG-Wisconsin Package Version 10, Madison, WI) programs "translate", "pilieup") (default values) and "prettybox".

Supplement 6.

(PPT 706 KB)

Southern Blots of 5 µg of genomic DNA from tobacco leaves, digested using XbaI SacI NdeI, HindIII, HaeII, EcorRI, and ClaI restriction enzymes. Digested DNA was separted in agarose gels, salt transferred to nylon membranes and probed using Tobacco preproprotein cDNA. Molecular markers anre indicated on the left (Kb). Und=undigested.

Supplement 7.

(PPT 187 KB)

Gel blot analyses of tobacco prosystemin mRNA in leaves of young tobacco plants exposed to either air or air containing methyl jasmonate vapors. Following methyl jasmonate treatment for 6 h, RNA extraction and gel blot analyses were performed as described in Bergey et al (1999).

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Pearce, G., Moura, D., Stratmann, J. et al. Production of multiple plant hormones from a single polyprotein precursor. Nature 411, 817–820 (2001). https://doi.org/10.1038/35081107

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