Small and Mighty: Peptide Hormones in Plant Biology

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Small and Mighty: Peptide Hormones in Plant Biology SMALL AND MIGHTY: PEPTIDE HORMONES IN PLANT BIOLOGY By Sonali Roy, Peter K. Lundquist, Michael K. Udvardi, and Wolf-Rüdiger Scheible © 2018 American Society of Plant Biologists Outline 1. Introduction to peptide hormones 2. Structure and generation of peptide hormones 3. Post-translational modifications and their presumed roles 4. Prediction and identification of peptide hormones 5. Mobility and perception of peptide hormones 6. Physiological roles of peptide hormones 7. The evolution of peptide hormones 8. Critical questions in peptide hormone field 9. Applications in biotechnology 10. Appendix: Peptide gene families © 2018 American Society of Plant Biologists PART 1 Introduction to peptide hormones © 2018 American Society of Plant Biologists What is a peptide hormone? • 5-60 amino acid residues • Serve as receptor ligands • Influence many developmental programs • Active at very low concentrations © 2018 American Society of Plant Biologists Peptide hormones control various developmental programs Shoot development Inflorescence development • Meristem maintenance • Inflorescence distribution • Cell division and expansion • Floral organ abscission • Plant height • Pollen tube germination, growth and • Biosynthesis of alkaloids guidance to ovule • Vascular differentiation • Stigma exudate deposition • Self-incompatability determinant Leaf development • Fruit shape • Stomatal patterning and distribution Seed and seed-pod development • Density of guard cells • Early and late embryo patterning • Leaf shape • Nutrient translocation in seeds • Leaf vascularization Response to biotic stresses Root development • Immune system and defense against Response to abiotic • Lateral root formation pathogens • Meristem maintenance • Associations with beneficial stresses • Gravitropism microorganisms • Nitrogen, phosphorus limitation • Root hair growth • Protection against herbivory • Stomatal control of transpiration • Casparian strip formation • Vascular differentiation © 2018 American Society of Plant Biologists Systemin was the first known plant peptide hormone ‘Isletin’ or insulin was the first known The first known plant peptide hormone peptide hormone was named systemin Wounding or insect 1889- Mering and Minkowski noted that 1972 T. Green and C. A boy with damage untreated removal of pancreas resulted in total diabetes A. Ryan noticed leaf severe in dogs. wounding caused diabetes. accumulation of 1921- Banting and Best hypothesized that proteinase inhibitors Islets of Langerhans in the pancreas in wounded and distal Fredrick produced an internal secretion that controlled unwounded leaves. Banting blood sugar levels. and Charles Concluded that a Best 1922- Extracted highly pure insulin and mobile factor treated patients (with J. Collips, J. Macleod). Inhibitors in systemically signals unwounded leaves the plant that it is under attack. 1926- John J. Abel crystallized insulin and found that it tested positive for the Biuret reaction, a test for proteins. 1991 Gregory Pearce HPLC chromatogram isolated the first 1951, 1952- Frederick Sanger identified Before & After insulin treatment peptide hormone the amino acid sequence of chain A (21 aa) and chain B (30aa) of the dipeptide • Called it systemin, Insulin. • 18 amino acid long • 14C labelled Structure of the insulin dipeptide 1980- Graeme I. Bell cloned the human synthetic systemin INSULIN gene commenting that it was mobile “consists of 3 exons and 2 introns • Active at 40 coding for a signal peptide, a B-chain, a Purify femtomolar! C-peptide, and an A-chain." Human INSULIN gene structure © 2018 American Society of Plant Biologists Peptide hormones share many characteristics of classical plant hormones Small molecule Peptide hormones hormones Similarities • Chemical Structure- • Regulate growth and • Chemical Structure- Derived from amino development even at minute Derived from acids (e.g., Indole-3- concentrations polypeptide chains acetic acid), isoprenoids (e.g., • Can act long distance Benzyl amino purine), • Biosynthesis- lipids (e.g., Methyl • Commonly require membrane Transcribed Jasmonate) ribosomally from a receptors for recognition single gene • Biosynthesis- End • Activity may require secondary products of a cellular metabolic pathway group modifications (e.g., hydroxylation) Y I Y T N SO4 SO4 Phytosulfokine peptide Auxin (IAA) © 2018 American Society of Plant Biologists Peptide hormones can act over short and long distances Cell-to-cell communication Organ-to-organ communication (Short distances) (Long distances) Example: In Legumes, Example: Yellow cells members of the CLE family indicate specific cells of the in roots signal to shoots root tip in which the peptide when they have enough CLE40 is expressed. nitrogen-fixing organs! SYMPLAST APOPLAST Image source: Root tip Modified from ?, Self © 2018 American Society of Plant Biologists Peptides can perform different functions But not all peptides are hormones! FREE BOUND PEPTIDE PEPTIDE 1. Can act as Signals ACTIVE COMPLEX e.g., CLE peptides RECEPTOR Co-RECEPTOR SIGNAL RELAY PLANT PLASMA MEMBRANE OUTSIDE 2. Can be anti-microbial INSIDE LEAKAGE OF IONS e.g., Defensins prevent infection BLOCK TRANSLATION MICROBIAL MEMBRANE PEPTIDE-BOND NO CLEAVAGE CLEAVAGE 3. Can inhibit peptidases Hasn’t been studied in plants to FUNCTIONAL PROTEIN COMPETETIVELY BINDS TO date! ACTIVE SITE INTRACELLULAR COMPARTMENTS © 2018 American Society of Plant Biologists PART 2 Structure and generation of peptide hormones © 2018 American Society of Plant Biologists Anatomy of a small signaling peptide Two major criteria are used to classify plant peptides (1) Primary sequence characteristics (2) Post-translational modifications N-TERMINAL VARIABLE REGION C-TERMINAL SIGNAL PEPTIDE SIGNALLING PEPTIDE • Directed to the • 5-60 amino acids long endoplasmic reticulum, • May contain different post- Golgi secretory pathway translational modifications © 2018 American Society of Plant Biologists Peptide hormones are classified into families Does the precursor itself YES YES perform a function? NO Peptide has a different Functional precursor function Does the peptide have YES FUNCTIONAL-PRECURSOR DERIVED PEPTIDES any post-translational modifications? (Tyrosine) Can be sulphated Is the peptide NO (Proline) Can be derived from a hydroxylated, POST-TRANSLATIONALLY single glycosylated Does the peptide MODIFIED PEPTIDES ribosomally YES have four or translated more cysteines? precursor? Di-sulphide bridges form between cysteines NO CYSTEINE-RICH PEPTIDES NONPRECURSOR DERIVED UNMODIFIED PEPTIDES NO 5’ region Pri- Short of mRNA miRNA ORF See Tavormina, P., De Coninck, B., Nikonorova, N., De Smet, I., and Cammue, B.P.A. (2015). The Plant Peptidome: An expanding repertoire of structural features and biological functions. Plant Cell 27: 2095–2118. © 2018 American Society of Plant Biologists Mature peptides are processed from longer precursors NUCLEUS ENDOPLASMIC RETICULUM GOLGI Pre-Pro-Peptide APPARATUS mRNA Peptide translation Pro-Peptide Pre-pro-peptide Precursor of pro-peptide Biologically active peptide Inactive peptide Image source: Modified from Wikipedia © 2018 American Society of Plant Biologists Peptide processing precedes peptide activity Signal-peptide peptidases Peptide cleavage occurs Release of signalling peptide remove N-terminal signals at specific motifs requires special enzymes Peptidase cleavage site (RxxL/RxLX) Peptidases (molecular scissors) Scissile Bond • Cleavage occurs at specific motifs. • C Score: Highest score at position immediately after the cleavage site • S Score: High score distinguishes proteins with signal peptide from those without • Peptidases/proteases/proteinases can belong to different families What do the D score and Y score depending on the amino acid indicate? Follow the link • Both upstream and downstream motifs residue that becomes nucleophilic http://www.cbs.dtu.dk/services/SignalP-4.1/ can contribute to site recognition. and cleaves the peptide bond. Stegmann, M., et al. , and Zipfel, C. (2017). The receptor kinase FER is a RALF-regulated scaffold controlling plant immune signaling. Science 355: 287–289; Schardon, K.,et tal. and Schaller, A. (2016). Precursor processing for plant peptide hormone maturation by subtilisin-like serine proteinases. Science 354: 1594–1597. © 2018 American Society of Plant Biologists Peptidases can be classified into different families Based on amino acid involved in the Based on position of cleavage site nucleophilic attack on the peptide bond N-terminus C-terminus N-terminus C-terminus Endopeptidase cleaves the non-terminal amino acids compared to Exopeptidases which hydrolyse terminal peptide bond to release a single amino acid. N-terminus C-terminus N-terminus C-terminus Exopeptidases can be further classified as Proteases can be classified into families based on sequence Aminopeptidases and Carboxypeptidases that homology and the amino acid residue that participates in the hydrolyse the peptide bond at the N-terminal and C- nucleophilic acid that cleaves the CONH peptide bond. terminal end respectively. Plant proteases can also be involved in protein turnover, degradation of misfolded proteins. Only a subset of the ~600 proteases in Arabidopsis are involved in the secretory pathway! © 2018 American Society of Plant Biologists Peptide activity requires proteases 1. Functional subtilisin serine protease is essential for peptide activity 35S-AtRALF23 35S-AtRALF23 OVEREXPRESS OVEREXPRESS Retarded root growth, site 1 protease Mutant Wild type like (no Wild type decreased hypocotyl phenotypic effect
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