Proteomics of nitrogen remobilization in poplar bark.

Seasonal nitrogen (N) cycling in temperate deciduous trees involves the accumulation of bark storage proteins (BSPs) in phloem parenchyma and xylem ray cells. BSPs are anabolized using recycled N during autumn leaf senescence and later become a source of N during spring shoot growth as they are catabolized. Little is known about the catabolic processes involved in remobilization and reutilization of N from BSPs in trees. In this study, we used multidimensional protein identification technology (MudPIT) and spectral counting to identify protein changes that occur in the bark during BSP catabolism. A total of 4,178 proteins were identified from bark prior to and during BSP catabolism. The majority (62%) of the proteins were found during BSP catabolism, indicating extensive remodeling of the proteome during renewed shoot growth and N remobilization. Among these proteins were 30 proteases, the relative abundances of which increased during BSP catabolism. These proteases spanned a range of families including members of the papain-like cysteine proteases, serine carboxypeptidases, and aspartyl proteases. These data identify, for the first time, candidate proteases that could potentially provide hydrolase activity required for N remobilization from BSPs and provide the foundation for research to advance our knowledge of poplar N cycling.

[1]  K. Müntz Protein dynamics and proteolysis in plant vacuoles. , 2007, Journal of experimental botany.

[2]  C. Ryan Protease Inhibitors in Plants: Genes for Improving Defenses Against Insects and Pathogens , 1990 .

[3]  K. Karrer,et al.  Two distinct gene subfamilies within the family of cysteine protease genes. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[4]  B Henrissat,et al.  Hydrophobic-cluster analysis of plant protein sequences. A domain homology between storage and lipid-transfer proteins. , 1988, The Biochemical journal.

[5]  B. Cooper,et al.  Proteomic pleiotropy of OpgGH, an operon necessary for efficient growth of Salmonella enterica serovar typhimurium under low-osmotic conditions. , 2012, Journal of proteome research.

[6]  D. Pantoja-Uceda,et al.  Solution structure of RicC3, a 2S albumin storage protein from Ricinus communis. , 2003, Biochemistry.

[7]  A. de Daruvar,et al.  Mapping the proteome of poplar and application to the discovery of drought‐stress responsive proteins , 2006, Proteomics.

[8]  Kanu Patel,et al.  An extracellular aspartic protease functions in Arabidopsis disease resistance signaling , 2004, The EMBO journal.

[9]  J. Sauter,et al.  Immunochemical Localization of a Storage Protein in Poplar Wood , 1988 .

[10]  G. Neilsen,et al.  Sources of N for leaf growth in a high-density apple (Malus domestica) orchard irrigated with ammonium nitrate solution. , 1997, Tree physiology.

[11]  J. Sauter,et al.  Biochemical, immunochemical, and ultrastructural studies of protein storage in poplar(Populus × canadensis ‘robusta’) wood , 1991, Planta.

[12]  H. Rennenberg,et al.  Thiol and amino acid composition of the xylem sap of poplar trees (Populus ×canadensis 'robusta') , 1994 .

[13]  J. Sauter,et al.  Seasonal variation of amino acids in the xylem sap of “Populus x canadensis” and its relation to protein body mobilization , 1992, Trees.

[14]  A. Schaller A cut above the rest: the regulatory function of plant proteases , 2004, Planta.

[15]  D. Luthe,et al.  A Unique 33-kD Cysteine Proteinase Accumulates in Response to Larval Feeding in Maize Genotypes Resistant to Fall Armyworm and Other Lepidoptera , 2000, Plant Cell.

[16]  Michael P Washburn,et al.  Proteomic analysis by multidimensional protein identification technology. , 2006, Methods in molecular biology.

[17]  M. Mann,et al.  Parts per Million Mass Accuracy on an Orbitrap Mass Spectrometer via Lock Mass Injection into a C-trap*S , 2005, Molecular & Cellular Proteomics.

[18]  D. Hedderley,et al.  Suppression of the cysteine protease, aleurain, delays floret senescence in Brassica oleracea , 2005, Plant Molecular Biology.

[19]  M. Bogyo,et al.  Subclassification and Biochemical Analysis of Plant Papain-Like Cysteine Proteases Displays Subfamily-Specific Characteristics1[C][W] , 2012, Plant Physiology.

[20]  K. Brown,et al.  Gene expression associated with N‐induced shifts in resource allocation in poplar , 2003 .

[21]  F. Bormann,et al.  Nutrient resorption in northern hardwood forests , 1982 .

[22]  B. Cooper,et al.  Nuclear proteomic changes linked to soybean rust resistance. , 2011, Molecular bioSystems.

[23]  F. Pontén,et al.  Correlations between RNA and protein expression profiles in 23 human cell lines , 2009, BMC Genomics.

[24]  I. Baldwin,et al.  Serine Protease Inhibitors Specifically Defend Solanum nigrum against Generalist Herbivores but Do Not Influence Plant Growth and Development[C][W] , 2010, Plant Cell.

[25]  Ben C. Stöver,et al.  TreeGraph 2: Combining and visualizing evidence from different phylogenetic analyses , 2010, BMC Bioinformatics.

[26]  N. Raikhel,et al.  The Plant Vacuolar Sorting Receptor Atelp Is Involved in Transport of Nh2-Terminal Propeptide-Containing Vacuolar Proteins in Arabidopsis thaliana , 2000, The Journal of cell biology.

[27]  F. Stuart Chapin,et al.  The Ecology and Economics of Storage in Plants , 1990 .

[28]  Elucidating the evolutionary history and expression patterns of nucleoside phosphorylase paralogs (vegetative storage proteins) in Populus and the plant kingdom , 2013, BMC Plant Biology.

[29]  C. Chapple,et al.  An Expression and Bioinformatics Analysis of the Arabidopsis Serine Carboxypeptidase-Like Gene Family1[w] , 2005, Plant Physiology.

[30]  P. A. Rea,et al.  The protein storage vacuole , 2001, The Journal of cell biology.

[31]  Paul Horton,et al.  Nucleic Acids Research Advance Access published May 21, 2007 WoLF PSORT: protein localization predictor , 2007 .

[32]  L. Fuchigami,et al.  Photoperiod control of poplar bark storage protein accumulation. , 1991, Plant physiology.

[33]  J. Yates,et al.  Large-scale analysis of the yeast proteome by multidimensional protein identification technology , 2001, Nature Biotechnology.

[34]  K. Chou,et al.  Plant-mPLoc: A Top-Down Strategy to Augment the Power for Predicting Plant Protein Subcellular Localization , 2010, PloS one.

[35]  C. Chapple,et al.  The sng2 mutant of Arabidopsis is defective in the gene encoding the serine carboxypeptidase-like protein sinapoylglucose:choline sinapoyltransferase. , 2001, The Plant journal : for cell and molecular biology.

[36]  I. Connerton,et al.  Recombinant pro-regions from papain and papaya proteinase IV-are selective high affinity inhibitors of the mature papaya enzymes. , 1995, Protein engineering.

[37]  Jörg Bohlmann,et al.  Analysis of the poplar phloem proteome and its response to leaf wounding. , 2009, Journal of proteome research.

[38]  J. Dyckmans,et al.  Influence of tree internal N status on uptake and translocation of C and N in beech: a dual 13C and 15N labeling approach. , 2001, Tree physiology.

[39]  G. Coleman,et al.  The poplar bark storage protein gene (Bspa) promoter is responsive to photoperiod and nitrogen in transgenic poplar and active in floral tissues, immature seeds and germinating seeds of transgenic tobacco , 2001, Plant Molecular Biology.

[40]  R. Boerner Foliar nutrient dynamics and nutrient use efficiency of four deciduous tree species in relation to site fertility , 1984 .

[41]  T. Morimoto,et al.  Tertiary and quaternary structures of 0.19 alpha-amylase inhibitor from wheat kernel determined by X-ray analysis at 2.06 A resolution. , 1997, Biochemistry.

[42]  S. Weinbaum,et al.  Quantitative estimates of uptake and internal cycling of (14)N-labeled fertilizer in mature walnut trees. , 1998, Tree physiology.

[43]  Koichiro Tamura,et al.  MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. , 2013, Molecular biology and evolution.

[44]  C Lehfeldt,et al.  Cloning of the SNG1 Gene of Arabidopsis Reveals a Role for a Serine Carboxypeptidase-like Protein as an Acyltransferase in Secondary Metabolism , 2000, Plant Cell.

[45]  A. Schlereth,et al.  Stored proteinases and the initiation of storage protein mobilization in seeds during germination and seedling growth. , 2001, Journal of experimental botany.

[46]  Neil D. Rawlings,et al.  MEROPS: the peptidase database , 2009, Nucleic Acids Res..

[47]  S. Brunak,et al.  Locating proteins in the cell using TargetP, SignalP and related tools , 2007, Nature Protocols.

[48]  Jonathan D. G. Jones,et al.  A Tomato Cysteine Protease Required for Cf-2-Dependent Disease Resistance and Suppression of Autonecrosis , 2002, Science.

[49]  Narmada Thanki,et al.  CDD: a Conserved Domain Database for the functional annotation of proteins , 2010, Nucleic Acids Res..

[50]  S. Wetzel,et al.  Spherical organelles, analogous to seed protein bodies, fluctuate seasonally in parenchymatous cells of hardwoods , 1989 .

[51]  Alex Bateman,et al.  MEROPS: the database of proteolytic enzymes, their substrates and inhibitors , 2011, Nucleic Acids Res..

[52]  A. Mattoo,et al.  Tomato Fruit Carboxypeptidase (Properties, Induction upon Wounding, and Immunocytochemical Localization) , 1996, Plant physiology.

[53]  H. Rennenberg,et al.  Seasonal nitrogen cycling in the bark of field-grown Grey poplar is correlated with meteorological factors and gene expression of bark storage proteins. , 2010, Tree physiology.

[54]  M. Tagliavini,et al.  Translocation of nitrogen in the xylem of field-grown cherry and poplar trees during remobilization. , 2006, Tree Physiology.

[55]  P. Lyu,et al.  Characterization and structural analyses of nonspecific lipid transfer protein 1 from mung bean. , 2005, Biochemistry.

[56]  G. Coleman,et al.  Evaluation of qPCR reference genes in two genotypes of Populus for use in photoperiod and low-temperature studies , 2012, BMC Research Notes.

[57]  H. Rennenberg,et al.  Perennial lifestyle--an adaptation to nutrient limitation? , 2010, Tree physiology.

[58]  C. Ryan,et al.  Characterization and localization of a wound-inducible type I serine-carboxypeptidase from leaves of tomato plants (Lycopersicon esculentum Mill.) , 2001, Planta.

[59]  P. Vitousek,et al.  Nitrogen Availability and Nitrogen Use Efficiency in Loblolly Pine Stands , 1986 .

[60]  Fanglian He,et al.  Protein storage vacuole acidification as a control of storage protein mobilization in soybeans. , 2007, Journal of experimental botany.

[61]  B. Cooper,et al.  Proteomic responses in Arabidopsis thaliana seedlings treated with ethylene. , 2011, Molecular bioSystems.

[62]  C. Ryan,et al.  Isolation and properties of carboxypeptidase from leaves of wounded tomato plants , 1980 .

[63]  R. Hoorn Plant proteases: From phenotypes to molecular mechanisms , 2008 .

[64]  F. Stuart Chapin,et al.  Seasonal Changes in Nitrogen and Phosphorus Fractions and Autumn Retranslocation in Evergreen and Deciduous Taiga Trees , 1983 .

[65]  L. Fuchigami,et al.  Physiological and Environmental Requirements for Poplar (Populus deltoides) Bark Storage Protein Degradation , 1993, Plant physiology.

[66]  J. Dunwell,et al.  Cupins: the most functionally diverse protein superfamily? , 2004, Phytochemistry.

[67]  P. Millard,et al.  Leaf demography and the seasonal internal cycling of nitrogen in sycamore (Acer pseudoplatanus L.) seedlings in relation to nitrogen supply , 1991 .

[68]  Scott A. Shaffer,et al.  Genetic Variation Shapes Protein Networks Mainly through Non-transcriptional Mechanisms , 2011, PLoS biology.

[69]  Jeffrey R. Whiteaker,et al.  Proteogenomic characterization of human colon and rectal cancer , 2014, Nature.

[70]  Martin Weih,et al.  Nitrogen storage and seasonal nitrogen cycling in Populus: bridging molecular physiology and ecophysiology. , 2005, The New phytologist.

[71]  N. Samatova,et al.  Detecting differential and correlated protein expression in label-free shotgun proteomics. , 2006, Journal of proteome research.

[72]  C. Plomion,et al.  Leaf proteome analysis of eight Populus ×euramericana genotypes: Genetic variation in drought response and in water‐use efficiency involves photosynthesis‐related proteins , 2009, Proteomics.

[73]  John D. Storey,et al.  Statistical significance for genomewide studies , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[74]  D. Naiman,et al.  Probability model for assessing proteins assembled from peptide sequences inferred from tandem mass spectrometry data. , 2007, Analytical chemistry.

[75]  P. Millard,et al.  Nitrogen storage and remobilization by trees: ecophysiological relevance in a changing world. , 2010, Tree physiology.

[76]  T. A. Valueva,et al.  Proteinase inhibitors and their function in plants: A review , 2005, Applied Biochemistry and Microbiology.

[77]  O. Franco,et al.  Plant storage proteins with antimicrobial activity: novel insights into plant defense mechanisms , 2011, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[78]  D. N. Perkins,et al.  Probability‐based protein identification by searching sequence databases using mass spectrometry data , 1999, Electrophoresis.

[79]  T. Mengiste,et al.  The Arabidopsis extracellular UNUSUAL SERINE PROTEASE INHIBITOR functions in resistance to necrotrophic fungi and insect herbivory. , 2011, The Plant journal : for cell and molecular biology.

[80]  Xiaochun Ge,et al.  An Arabidopsis aspartic protease functions as an anti‐cell‐death component in reproduction and embryogenesis , 2005, EMBO reports.

[81]  R. Jung,et al.  The Proteolytic Processing of Seed Storage Proteins in Arabidopsis Embryo Cells Starts in the Multivesicular Bodies[W] , 2006, The Plant Cell Online.

[82]  M. Nishimura,et al.  A slow maturation of a cysteine protease with a granulin domain in the vacuoles of senescing Arabidopsis leaves. , 2001, Plant physiology.

[83]  D. Naiman,et al.  Quantitative Proteomic Analysis of Bean Plants Infected by a Virulent and Avirulent Obligate Rust Fungus *S , 2009, Molecular & Cellular Proteomics.

[84]  J. Yates,et al.  An automated multidimensional protein identification technology for shotgun proteomics. , 2001, Analytical chemistry.

[85]  C. P. Constabel,et al.  Proteomic analysis of hybrid poplar xylem sap. , 2009, Phytochemistry.

[86]  G. Loake,et al.  Involvement of cathepsin B in the plant disease resistance hypersensitive response. , 2007, The Plant journal : for cell and molecular biology.

[87]  K. Wilson,et al.  Refined crystal structures of subtilisin novo in complex with wild-type and two mutant eglins. Comparison with other serine proteinase inhibitor complexes. , 1991, Journal of molecular biology.

[88]  Zhou Du,et al.  agriGO: a GO analysis toolkit for the agricultural community , 2010, Nucleic Acids Res..

[89]  Peter M. Vitousek,et al.  Nutrient Cycling and Nutrient Use Efficiency , 1982, The American Naturalist.

[90]  R. Boerner,et al.  Foliar nutrient dynamics and nutrient use efficiency in Cornus florida , 1985, Oecologia.

[91]  L. Staehelin,et al.  Protein Storage Vacuoles Are Transformed into Lytic Vacuoles in Root Meristematic Cells of Germinating Seedlings by Multiple, Cell Type-Specific Mechanisms1[W] , 2011, Plant Physiology.

[92]  B. Cooper,et al.  Relative, label-free protein quantitation: Spectral counting error statistics from nine replicate MudPIT samples , 2010, Journal of the American Society for Mass Spectrometry.

[93]  E. Marcotte,et al.  Global signatures of protein and mRNA expression levelsw , 2009 .

[94]  J. Kader LIPID-TRANSFER PROTEINS IN PLANTS. , 1996, Annual review of plant physiology and plant molecular biology.

[95]  P. Staswick Storage Proteins of Vegetative Plant Tissues , 1994 .

[96]  K. Apel,et al.  Protein bodies in ray cells of Populus x canadensis Moench ‘robusta’ , 2004, Planta.

[97]  Gene Kwan,et al.  Role of the intramolecular hydrogen bond network in the inhibitory power of chymotrypsin inhibitor 2. , 2005, Biochemistry.

[98]  G. Coleman,et al.  Phytochrome-mediated photoperiod perception, shoot growth, glutamine, calcium, and protein phosphorylation influence the activity of the poplar bark storage protein gene promoter (bspA). , 2001, Plant physiology.

[99]  H. Bäumlein,et al.  Storage and mobilization as antagonistic functional constraints on seed storage globulin evolution. , 2003, Journal of experimental botany.

[100]  Dong Wook Lee,et al.  Trafficking of Vacuolar Proteins: The Crucial Role of Arabidopsis Vacuolar Protein Sorting 29 in Recycling Vacuolar Sorting Receptor[W] , 2012, Plant Cell.

[101]  A. Schlereth,et al.  Differential tissue-specific expression of cysteine proteinases forms the basis for the fine-tuned mobilization of storage globulin during and after germination in legume seeds , 2001, Planta.

[102]  F. Dal Degan,et al.  The expression of serine carboxypeptidases during maturation and germination of the barley grain. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[103]  R. Aerts Nutrient use efficiency in evergreen and deciduous species from heathlands , 1990, Oecologia.

[104]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[105]  C. Funk,et al.  Protease gene families in Populus and Arabidopsis , 2006, BMC Plant Biology.

[106]  THH. Chen,et al.  Poplar Bark Storage Protein and a Related Wound-Induced Gene Are Differentially Induced by Nitrogen , 1994, Plant physiology.