Isolation and characterization of galactinol synthases from hybrid poplar.
Identifieur interne : 002B93 ( Main/Corpus ); précédent : 002B92; suivant : 002B94Isolation and characterization of galactinol synthases from hybrid poplar.
Auteurs : Faride Unda ; Thomas Canam ; Lindsay Preston ; Shawn D. MansfieldSource :
- Journal of experimental botany [ 1460-2431 ] ; 2012.
English descriptors
- KwdEn :
- Amino Acid Sequence (MeSH), Chimera (MeSH), Conserved Sequence (MeSH), DNA, Complementary (genetics), Galactosyltransferases (genetics), Galactosyltransferases (metabolism), Molecular Sequence Data (MeSH), Oligosaccharides (metabolism), Phloem (metabolism), Phylogeny (MeSH), Pichia (genetics), Plant Leaves (genetics), Plant Leaves (metabolism), Plant Proteins (genetics), Plant Proteins (metabolism), Populus (enzymology), Populus (genetics), Populus (metabolism), Protein Isoforms (MeSH), Raffinose (metabolism), Sequence Alignment (MeSH), Stress, Physiological (MeSH), Trees (MeSH).
- MESH :
- chemical , genetics : DNA, Complementary, Galactosyltransferases, Plant Proteins.
- chemical , metabolism : Galactosyltransferases, Oligosaccharides, Plant Proteins, Raffinose.
- enzymology : Populus.
- genetics : Pichia, Plant Leaves, Populus.
- metabolism : Phloem, Plant Leaves, Populus.
- Amino Acid Sequence, Chimera, Conserved Sequence, Molecular Sequence Data, Phylogeny, Protein Isoforms, Sequence Alignment, Stress, Physiological, Trees.
Abstract
The raffinose family of oligosaccharides (RFOs) serve as transport carbohydrates in the phloem, storage compounds in sink tissues, and putative biological agents to combat both abiotic and biotic stress in several plant species. To investigate further the functional roles of this class of compounds in trees, two cDNAs encoding galactinol synthase (GolS, EC 2.4.1.123), which catalyses the first step in the biosynthesis of RFOs, were identified and cloned from hybrid poplar (Populus alba×grandidentata). Phylogenetic analyses of the Populus GolS isoforms with other known GolS proteins suggested a putative role for these enzymes during biotic or abiotic stress in hybrid poplar. The predicted protein sequences of both isoforms (Pa×gGolSI and Pa×gGolSII) showed characteristics of GolS proteins from other species, including a serine phosphorylation site and the ASAAP pentapeptide hydrophobic domain. Kinetic analyses of recombinant Pa×gGolSI and Pa×gGolSII resulted in K(m) values for UPD-galactose of 0.80 and 0.65 mM and V(max) values of 657.5 and 1245 nM min(-1), respectively. Pa×gGolSI inherently possessed a broader pH and temperature range when compared with Pa×gGolSII. Interestingly, spatial and temporal expression analyses revealed that Pa×gGolSII transcript levels varied seasonally, while Pa×gGolSI did not, implying temperature-regulated transcriptional control of this gene in addition to the observed thermosensitivity of the respective enzyme. This evidence suggested that Pa×gGolSI may be involved in basic metabolic activities such as storage, while Pa×gGolSII is probably involved in seasonal mobilization of carbohydrates.
DOI: 10.1093/jxb/err411
PubMed: 22197892
PubMed Central: PMC3295395
Links to Exploration step
pubmed:22197892Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Isolation and characterization of galactinol synthases from hybrid poplar.</title><author><name sortKey="Unda, Faride" sort="Unda, Faride" uniqKey="Unda F" first="Faride" last="Unda">Faride Unda</name><affiliation><nlm:affiliation>Department of Wood Science, The University of British Columbia, Vancouver, BC, Canada.</nlm:affiliation></affiliation></author><author><name sortKey="Canam, Thomas" sort="Canam, Thomas" uniqKey="Canam T" first="Thomas" last="Canam">Thomas Canam</name></author><author><name sortKey="Preston, Lindsay" sort="Preston, Lindsay" uniqKey="Preston L" first="Lindsay" last="Preston">Lindsay Preston</name></author><author><name sortKey="Mansfield, Shawn D" sort="Mansfield, Shawn D" uniqKey="Mansfield S" first="Shawn D" last="Mansfield">Shawn D. Mansfield</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2012">2012</date><idno type="RBID">pubmed:22197892</idno><idno type="pmid">22197892</idno><idno type="doi">10.1093/jxb/err411</idno><idno type="pmc">PMC3295395</idno><idno type="wicri:Area/Main/Corpus">002B93</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">002B93</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Isolation and characterization of galactinol synthases from hybrid poplar.</title><author><name sortKey="Unda, Faride" sort="Unda, Faride" uniqKey="Unda F" first="Faride" last="Unda">Faride Unda</name><affiliation><nlm:affiliation>Department of Wood Science, The University of British Columbia, Vancouver, BC, Canada.</nlm:affiliation></affiliation></author><author><name sortKey="Canam, Thomas" sort="Canam, Thomas" uniqKey="Canam T" first="Thomas" last="Canam">Thomas Canam</name></author><author><name sortKey="Preston, Lindsay" sort="Preston, Lindsay" uniqKey="Preston L" first="Lindsay" last="Preston">Lindsay Preston</name></author><author><name sortKey="Mansfield, Shawn D" sort="Mansfield, Shawn D" uniqKey="Mansfield S" first="Shawn D" last="Mansfield">Shawn D. Mansfield</name></author></analytic><series><title level="j">Journal of experimental botany</title><idno type="eISSN">1460-2431</idno><imprint><date when="2012" type="published">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence (MeSH)</term><term>Chimera (MeSH)</term><term>Conserved Sequence (MeSH)</term><term>DNA, Complementary (genetics)</term><term>Galactosyltransferases (genetics)</term><term>Galactosyltransferases (metabolism)</term><term>Molecular Sequence Data (MeSH)</term><term>Oligosaccharides (metabolism)</term><term>Phloem (metabolism)</term><term>Phylogeny (MeSH)</term><term>Pichia (genetics)</term><term>Plant Leaves (genetics)</term><term>Plant Leaves (metabolism)</term><term>Plant Proteins (genetics)</term><term>Plant Proteins (metabolism)</term><term>Populus (enzymology)</term><term>Populus (genetics)</term><term>Populus (metabolism)</term><term>Protein Isoforms (MeSH)</term><term>Raffinose (metabolism)</term><term>Sequence Alignment (MeSH)</term><term>Stress, Physiological (MeSH)</term><term>Trees (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Complementary</term><term>Galactosyltransferases</term><term>Plant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Galactosyltransferases</term><term>Oligosaccharides</term><term>Plant Proteins</term><term>Raffinose</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Populus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Pichia</term><term>Plant Leaves</term><term>Populus</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Phloem</term><term>Plant Leaves</term><term>Populus</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Chimera</term><term>Conserved Sequence</term><term>Molecular Sequence Data</term><term>Phylogeny</term><term>Protein Isoforms</term><term>Sequence Alignment</term><term>Stress, Physiological</term><term>Trees</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The raffinose family of oligosaccharides (RFOs) serve as transport carbohydrates in the phloem, storage compounds in sink tissues, and putative biological agents to combat both abiotic and biotic stress in several plant species. To investigate further the functional roles of this class of compounds in trees, two cDNAs encoding galactinol synthase (GolS, EC 2.4.1.123), which catalyses the first step in the biosynthesis of RFOs, were identified and cloned from hybrid poplar (Populus alba×grandidentata). Phylogenetic analyses of the Populus GolS isoforms with other known GolS proteins suggested a putative role for these enzymes during biotic or abiotic stress in hybrid poplar. The predicted protein sequences of both isoforms (Pa×gGolSI and Pa×gGolSII) showed characteristics of GolS proteins from other species, including a serine phosphorylation site and the ASAAP pentapeptide hydrophobic domain. Kinetic analyses of recombinant Pa×gGolSI and Pa×gGolSII resulted in K(m) values for UPD-galactose of 0.80 and 0.65 mM and V(max) values of 657.5 and 1245 nM min(-1), respectively. Pa×gGolSI inherently possessed a broader pH and temperature range when compared with Pa×gGolSII. Interestingly, spatial and temporal expression analyses revealed that Pa×gGolSII transcript levels varied seasonally, while Pa×gGolSI did not, implying temperature-regulated transcriptional control of this gene in addition to the observed thermosensitivity of the respective enzyme. This evidence suggested that Pa×gGolSI may be involved in basic metabolic activities such as storage, while Pa×gGolSII is probably involved in seasonal mobilization of carbohydrates.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">22197892</PMID><DateCompleted><Year>2014</Year><Month>06</Month><Day>04</Day></DateCompleted><DateRevised><Year>2018</Year><Month>12</Month><Day>19</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1460-2431</ISSN><JournalIssue CitedMedium="Internet"><Volume>63</Volume><Issue>5</Issue><PubDate><Year>2012</Year><Month>Mar</Month></PubDate></JournalIssue><Title>Journal of experimental botany</Title><ISOAbbreviation>J Exp Bot</ISOAbbreviation></Journal><ArticleTitle>Isolation and characterization of galactinol synthases from hybrid poplar.</ArticleTitle><Pagination><MedlinePgn>2059-69</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1093/jxb/err411</ELocationID><Abstract><AbstractText>The raffinose family of oligosaccharides (RFOs) serve as transport carbohydrates in the phloem, storage compounds in sink tissues, and putative biological agents to combat both abiotic and biotic stress in several plant species. To investigate further the functional roles of this class of compounds in trees, two cDNAs encoding galactinol synthase (GolS, EC 2.4.1.123), which catalyses the first step in the biosynthesis of RFOs, were identified and cloned from hybrid poplar (Populus alba×grandidentata). Phylogenetic analyses of the Populus GolS isoforms with other known GolS proteins suggested a putative role for these enzymes during biotic or abiotic stress in hybrid poplar. The predicted protein sequences of both isoforms (Pa×gGolSI and Pa×gGolSII) showed characteristics of GolS proteins from other species, including a serine phosphorylation site and the ASAAP pentapeptide hydrophobic domain. Kinetic analyses of recombinant Pa×gGolSI and Pa×gGolSII resulted in K(m) values for UPD-galactose of 0.80 and 0.65 mM and V(max) values of 657.5 and 1245 nM min(-1), respectively. Pa×gGolSI inherently possessed a broader pH and temperature range when compared with Pa×gGolSII. Interestingly, spatial and temporal expression analyses revealed that Pa×gGolSII transcript levels varied seasonally, while Pa×gGolSI did not, implying temperature-regulated transcriptional control of this gene in addition to the observed thermosensitivity of the respective enzyme. This evidence suggested that Pa×gGolSI may be involved in basic metabolic activities such as storage, while Pa×gGolSII is probably involved in seasonal mobilization of carbohydrates.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Unda</LastName><ForeName>Faride</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>Department of Wood Science, The University of British Columbia, Vancouver, BC, Canada.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Canam</LastName><ForeName>Thomas</ForeName><Initials>T</Initials></Author><Author ValidYN="Y"><LastName>Preston</LastName><ForeName>Lindsay</ForeName><Initials>L</Initials></Author><Author ValidYN="Y"><LastName>Mansfield</LastName><ForeName>Shawn D</ForeName><Initials>SD</Initials></Author></AuthorList><Language>eng</Language><DataBankList 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