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Cloning and endogenous expression of a Eucalyptus grandis UDP-glucose dehydrogenase cDNA
UDP-glucose dehydrogenase (UGDH) catalyzes the oxidation of UDP-glucose (UDP-Glc) to UDP-glucuronate (UDP-GlcA), a key sugar nucleotide involved in the biosynthesis of plant cell wall polysaccharides. A full-length cDNA fragment coding for UGDH was cloned from the cambial region of 6-month-old E. gr...
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Published in: | Genetics and molecular biology 2010-01, Vol.33 (4), p.686-695 |
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creator | Labate, Mônica T Veneziano Bertolo, Ana L Ferreira do Nascimento, Daniela Defávari Gutmanis, Gunta de Andrade, Alexander Rodrigues, Maria J Calderan Camargo, Eduardo L O Boaretto, Luis Felipe Moon, David H Bragatto, Juliano Labate, Carlos A |
description | UDP-glucose dehydrogenase (UGDH) catalyzes the oxidation of UDP-glucose (UDP-Glc) to UDP-glucuronate (UDP-GlcA), a key sugar nucleotide involved in the biosynthesis of plant cell wall polysaccharides. A full-length cDNA fragment coding for UGDH was cloned from the cambial region of 6-month-old E. grandis saplings by RT-PCR. The 1443-bp-ORF encodes a protein of 480 amino acids with a predicted molecular weight of 53 kDa. The recombinant protein expressed in Escherichia coli catalyzed the conversion of UDP-Glc to UDP-GlcA, confirming that the cloned cDNA encodes UGDH. The deduced amino acid sequence of the cDNA showed a high degree of identity with UGDH from several plant species. The Southern blot assay indicated that more than one copy of UGDH is present in Eucalyptus. These results were also confirmed by the proteomic analysis of the cambial region of 3- and 22-year-old E. grandis trees by 2-DE and LC-MS/MS, showing that at least two isoforms are present. The cloned gene is mainly expressed in roots, stem and bark of 6-month-old saplings, with a lower expression in leaves. High expression levels were also observed in the cambial region of 3- and 22-year-old trees. The results described in this paper provide a further view of the hemicellulose biosynthesis during wood formation in E. grandis. |
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A full-length cDNA fragment coding for UGDH was cloned from the cambial region of 6-month-old E. grandis saplings by RT-PCR. The 1443-bp-ORF encodes a protein of 480 amino acids with a predicted molecular weight of 53 kDa. The recombinant protein expressed in Escherichia coli catalyzed the conversion of UDP-Glc to UDP-GlcA, confirming that the cloned cDNA encodes UGDH. The deduced amino acid sequence of the cDNA showed a high degree of identity with UGDH from several plant species. The Southern blot assay indicated that more than one copy of UGDH is present in Eucalyptus. These results were also confirmed by the proteomic analysis of the cambial region of 3- and 22-year-old E. grandis trees by 2-DE and LC-MS/MS, showing that at least two isoforms are present. The cloned gene is mainly expressed in roots, stem and bark of 6-month-old saplings, with a lower expression in leaves. High expression levels were also observed in the cambial region of 3- and 22-year-old trees. The results described in this paper provide a further view of the hemicellulose biosynthesis during wood formation in E. grandis.</description><identifier>ISSN: 1415-4757</identifier><identifier>ISSN: 1678-4685</identifier><identifier>EISSN: 1678-4685</identifier><identifier>DOI: 10.1590/S1415-47572010005000078</identifier><identifier>PMID: 21637578</identifier><language>eng</language><publisher>Brazil: Sociedade Brasileira de Genética</publisher><subject>BIOCHEMISTRY & MOLECULAR BIOLOGY ; cell wall ; GENETICS & HEREDITY ; hemicellulose ; pectin ; Plant Genetics ; UDP-glucose ; UDP-glucuronate</subject><ispartof>Genetics and molecular biology, 2010-01, Vol.33 (4), p.686-695</ispartof><rights>Copyright © 2010, Sociedade Brasileira de Genética. 2010</rights><rights>This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c521t-48cfc5c3293744b8978edc57d9776f69958935357da87e907f29ae7d441cc3d53</citedby><cites>FETCH-LOGICAL-c521t-48cfc5c3293744b8978edc57d9776f69958935357da87e907f29ae7d441cc3d53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,864,885,2102,24150,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21637578$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Labate, Mônica T Veneziano</creatorcontrib><creatorcontrib>Bertolo, Ana L Ferreira</creatorcontrib><creatorcontrib>do Nascimento, Daniela Defávari</creatorcontrib><creatorcontrib>Gutmanis, Gunta</creatorcontrib><creatorcontrib>de Andrade, Alexander</creatorcontrib><creatorcontrib>Rodrigues, Maria J Calderan</creatorcontrib><creatorcontrib>Camargo, Eduardo L O</creatorcontrib><creatorcontrib>Boaretto, Luis Felipe</creatorcontrib><creatorcontrib>Moon, David H</creatorcontrib><creatorcontrib>Bragatto, Juliano</creatorcontrib><creatorcontrib>Labate, Carlos A</creatorcontrib><title>Cloning and endogenous expression of a Eucalyptus grandis UDP-glucose dehydrogenase cDNA</title><title>Genetics and molecular biology</title><addtitle>Genet Mol Biol</addtitle><description>UDP-glucose dehydrogenase (UGDH) catalyzes the oxidation of UDP-glucose (UDP-Glc) to UDP-glucuronate (UDP-GlcA), a key sugar nucleotide involved in the biosynthesis of plant cell wall polysaccharides. A full-length cDNA fragment coding for UGDH was cloned from the cambial region of 6-month-old E. grandis saplings by RT-PCR. The 1443-bp-ORF encodes a protein of 480 amino acids with a predicted molecular weight of 53 kDa. The recombinant protein expressed in Escherichia coli catalyzed the conversion of UDP-Glc to UDP-GlcA, confirming that the cloned cDNA encodes UGDH. The deduced amino acid sequence of the cDNA showed a high degree of identity with UGDH from several plant species. The Southern blot assay indicated that more than one copy of UGDH is present in Eucalyptus. These results were also confirmed by the proteomic analysis of the cambial region of 3- and 22-year-old E. grandis trees by 2-DE and LC-MS/MS, showing that at least two isoforms are present. The cloned gene is mainly expressed in roots, stem and bark of 6-month-old saplings, with a lower expression in leaves. High expression levels were also observed in the cambial region of 3- and 22-year-old trees. The results described in this paper provide a further view of the hemicellulose biosynthesis during wood formation in E. grandis.</description><subject>BIOCHEMISTRY & MOLECULAR BIOLOGY</subject><subject>cell wall</subject><subject>GENETICS & HEREDITY</subject><subject>hemicellulose</subject><subject>pectin</subject><subject>Plant Genetics</subject><subject>UDP-glucose</subject><subject>UDP-glucuronate</subject><issn>1415-4757</issn><issn>1678-4685</issn><issn>1678-4685</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>DOA</sourceid><recordid>eNplUl1rVDEQDaLYuvoX9L75dGu-P16Esq1aKCpowbeQTXJvs2STNblX3H9vtrutFgNDJjNzzhwmA8AbBM8QU_DdN0QR66lgAkMEIWTNoJBPwCniQvaUS_a0-fdFJ-BFrWsIsSAMPwcnGHHSwvIU_FjGnEIaO5Nc55PLo095rp3_vS2-1pBTl4fOdJezNXG3nVpqLK021O7m4ms_xtnm6jvnb3eu7MGmvezF5_OX4NlgYvWvjvcC3Hy4_L781F9_-Xi1PL_uLcNo6qm0g2WWYEUEpSuphPTOMuGUEHzgSjGpCCMtYKTwCooBK-OFoxRZSxwjC3B14HXZrPW2hI0pO51N0HeBXEZtyhRs9JpwyhGzTiDrKeNUYm4h5sIwStxqsI3r7MBVbfAx63WeS2ri9d249d9xQ9oM7Zu_PwC282rTdPs0FRMfqXicSeFWj_mXJpA0KagRvD0SlPxz9nXSm1Ctj9Ek375BSwGx4qSdBRCHSltyrcUPD10Q1Pud-E_kcSca8vW_Ih9w90tA_gCfH69Z</recordid><startdate>20100101</startdate><enddate>20100101</enddate><creator>Labate, Mônica T Veneziano</creator><creator>Bertolo, Ana L Ferreira</creator><creator>do Nascimento, Daniela Defávari</creator><creator>Gutmanis, Gunta</creator><creator>de Andrade, Alexander</creator><creator>Rodrigues, Maria J Calderan</creator><creator>Camargo, Eduardo L O</creator><creator>Boaretto, Luis Felipe</creator><creator>Moon, David H</creator><creator>Bragatto, Juliano</creator><creator>Labate, Carlos A</creator><general>Sociedade Brasileira de Genética</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope><scope>GPN</scope><scope>DOA</scope></search><sort><creationdate>20100101</creationdate><title>Cloning and endogenous expression of a Eucalyptus grandis UDP-glucose dehydrogenase cDNA</title><author>Labate, Mônica T Veneziano ; Bertolo, Ana L Ferreira ; do Nascimento, Daniela Defávari ; Gutmanis, Gunta ; de Andrade, Alexander ; Rodrigues, Maria J Calderan ; Camargo, Eduardo L O ; Boaretto, Luis Felipe ; Moon, David H ; Bragatto, Juliano ; Labate, Carlos A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c521t-48cfc5c3293744b8978edc57d9776f69958935357da87e907f29ae7d441cc3d53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>BIOCHEMISTRY & MOLECULAR BIOLOGY</topic><topic>cell wall</topic><topic>GENETICS & HEREDITY</topic><topic>hemicellulose</topic><topic>pectin</topic><topic>Plant Genetics</topic><topic>UDP-glucose</topic><topic>UDP-glucuronate</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Labate, Mônica T Veneziano</creatorcontrib><creatorcontrib>Bertolo, Ana L Ferreira</creatorcontrib><creatorcontrib>do Nascimento, Daniela Defávari</creatorcontrib><creatorcontrib>Gutmanis, Gunta</creatorcontrib><creatorcontrib>de Andrade, Alexander</creatorcontrib><creatorcontrib>Rodrigues, Maria J Calderan</creatorcontrib><creatorcontrib>Camargo, Eduardo L O</creatorcontrib><creatorcontrib>Boaretto, Luis Felipe</creatorcontrib><creatorcontrib>Moon, David H</creatorcontrib><creatorcontrib>Bragatto, Juliano</creatorcontrib><creatorcontrib>Labate, Carlos A</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>SciELO</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>Genetics and molecular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Labate, Mônica T Veneziano</au><au>Bertolo, Ana L Ferreira</au><au>do Nascimento, Daniela Defávari</au><au>Gutmanis, Gunta</au><au>de Andrade, Alexander</au><au>Rodrigues, Maria J Calderan</au><au>Camargo, Eduardo L O</au><au>Boaretto, Luis Felipe</au><au>Moon, David H</au><au>Bragatto, Juliano</au><au>Labate, Carlos A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning and endogenous expression of a Eucalyptus grandis UDP-glucose dehydrogenase cDNA</atitle><jtitle>Genetics and molecular biology</jtitle><addtitle>Genet Mol Biol</addtitle><date>2010-01-01</date><risdate>2010</risdate><volume>33</volume><issue>4</issue><spage>686</spage><epage>695</epage><pages>686-695</pages><issn>1415-4757</issn><issn>1678-4685</issn><eissn>1678-4685</eissn><abstract>UDP-glucose dehydrogenase (UGDH) catalyzes the oxidation of UDP-glucose (UDP-Glc) to UDP-glucuronate (UDP-GlcA), a key sugar nucleotide involved in the biosynthesis of plant cell wall polysaccharides. A full-length cDNA fragment coding for UGDH was cloned from the cambial region of 6-month-old E. grandis saplings by RT-PCR. The 1443-bp-ORF encodes a protein of 480 amino acids with a predicted molecular weight of 53 kDa. The recombinant protein expressed in Escherichia coli catalyzed the conversion of UDP-Glc to UDP-GlcA, confirming that the cloned cDNA encodes UGDH. The deduced amino acid sequence of the cDNA showed a high degree of identity with UGDH from several plant species. The Southern blot assay indicated that more than one copy of UGDH is present in Eucalyptus. These results were also confirmed by the proteomic analysis of the cambial region of 3- and 22-year-old E. grandis trees by 2-DE and LC-MS/MS, showing that at least two isoforms are present. The cloned gene is mainly expressed in roots, stem and bark of 6-month-old saplings, with a lower expression in leaves. High expression levels were also observed in the cambial region of 3- and 22-year-old trees. 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subjects | BIOCHEMISTRY & MOLECULAR BIOLOGY cell wall GENETICS & HEREDITY hemicellulose pectin Plant Genetics UDP-glucose UDP-glucuronate |
title | Cloning and endogenous expression of a Eucalyptus grandis UDP-glucose dehydrogenase cDNA |
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