An in vivo internal deletion in the N‐terminus region of Arabidopsis cystathionine γ‐synthase results in CGS expression that is insensitive to methionine

Summary Cystathionine γ‐synthase (CGS), the first enzyme of methionine biosynthesis in higher plants, plays an important role in the biosynthesis pathway and in regulating methionine metabolism in plant cells. In response to methionine, the expression of this enzyme is regulated via amino acid seque...

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Veröffentlicht in:The Plant journal : for cell and molecular biology 2006-03, Vol.45 (6), p.955-967
Hauptverfasser: Hacham, Yael, Schuster, Gadi, Amir, Rachel
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Amir, Rachel
description Summary Cystathionine γ‐synthase (CGS), the first enzyme of methionine biosynthesis in higher plants, plays an important role in the biosynthesis pathway and in regulating methionine metabolism in plant cells. In response to methionine, the expression of this enzyme is regulated via amino acid sequences located in its N‐terminal. Here, using reverse transcription PCR and ribonuclease protection analysis, we demonstrate that, in addition to the full‐length CGS transcript, a deleted form exists in Arabidopsis. The deleted transcript of CGS that lacks 90 or 87 nt located internally in the regulatory N‐terminal region of CGS maintains the reading frame of the protein. Its association with polyribosomes indicates that this deleted form of CGS is translated. In order to study the function of this deleted form of CGS, we overexpressed it in transgenic tobacco plants. We found that the transgenic plants engineered to express only the deleted form of CGS accumulated methionine to a much higher level than those that expressed the full‐length CGS. Furthermore, in vitro feeding experiments revealed that the deleted form of CGS is not subject to feedback regulation by methionine, as reported for the full‐length transcript. Therefore, although most likely produced from the full‐length CGS, the transcript of the deleted form is insensitive to methionine application and its expression may be important for maintaining methionine metabolism even in the presence of a high level of methionine.
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In response to methionine, the expression of this enzyme is regulated via amino acid sequences located in its N‐terminal. Here, using reverse transcription PCR and ribonuclease protection analysis, we demonstrate that, in addition to the full‐length CGS transcript, a deleted form exists in Arabidopsis. The deleted transcript of CGS that lacks 90 or 87 nt located internally in the regulatory N‐terminal region of CGS maintains the reading frame of the protein. Its association with polyribosomes indicates that this deleted form of CGS is translated. In order to study the function of this deleted form of CGS, we overexpressed it in transgenic tobacco plants. We found that the transgenic plants engineered to express only the deleted form of CGS accumulated methionine to a much higher level than those that expressed the full‐length CGS. Furthermore, in vitro feeding experiments revealed that the deleted form of CGS is not subject to feedback regulation by methionine, as reported for the full‐length transcript. 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Physicochemical requirements ; Methionine - metabolism ; Methionine - physiology ; methionine synthesis ; Models, Biological ; Molecular Sequence Data ; mRNA stability ; Nicotiana - genetics ; Plant physiology and development ; Plants, Genetically Modified - genetics ; Plants, Genetically Modified - metabolism ; Polyribosomes - metabolism ; Protein Structure, Tertiary ; RNA Stability ; RNA, Messenger - metabolism ; Sequence Deletion ; sulfur amino acids ; transgenic plants</subject><ispartof>The Plant journal : for cell and molecular biology, 2006-03, Vol.45 (6), p.955-967</ispartof><rights>2006 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4281-7d049d79e98637cc4fb41241e5c8bdb1ed7a02eeb444307bcd78741bd4e986733</citedby><cites>FETCH-LOGICAL-c4281-7d049d79e98637cc4fb41241e5c8bdb1ed7a02eeb444307bcd78741bd4e986733</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1365-313X.2006.02661.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1365-313X.2006.02661.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,1427,27901,27902,45550,45551,46384,46808</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=17569525$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16507086$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hacham, Yael</creatorcontrib><creatorcontrib>Schuster, Gadi</creatorcontrib><creatorcontrib>Amir, Rachel</creatorcontrib><title>An in vivo internal deletion in the N‐terminus region of Arabidopsis cystathionine γ‐synthase results in CGS expression that is insensitive to methionine</title><title>The Plant journal : for cell and molecular biology</title><addtitle>Plant J</addtitle><description>Summary Cystathionine γ‐synthase (CGS), the first enzyme of methionine biosynthesis in higher plants, plays an important role in the biosynthesis pathway and in regulating methionine metabolism in plant cells. In response to methionine, the expression of this enzyme is regulated via amino acid sequences located in its N‐terminal. Here, using reverse transcription PCR and ribonuclease protection analysis, we demonstrate that, in addition to the full‐length CGS transcript, a deleted form exists in Arabidopsis. The deleted transcript of CGS that lacks 90 or 87 nt located internally in the regulatory N‐terminal region of CGS maintains the reading frame of the protein. Its association with polyribosomes indicates that this deleted form of CGS is translated. In order to study the function of this deleted form of CGS, we overexpressed it in transgenic tobacco plants. We found that the transgenic plants engineered to express only the deleted form of CGS accumulated methionine to a much higher level than those that expressed the full‐length CGS. Furthermore, in vitro feeding experiments revealed that the deleted form of CGS is not subject to feedback regulation by methionine, as reported for the full‐length transcript. Therefore, although most likely produced from the full‐length CGS, the transcript of the deleted form is insensitive to methionine application and its expression may be important for maintaining methionine metabolism even in the presence of a high level of methionine.</description><subject>Amino Acid Sequence</subject><subject>Arabidopsis</subject><subject>Arabidopsis - enzymology</subject><subject>Arabidopsis - genetics</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Carbon-Oxygen Lyases - chemistry</subject><subject>Carbon-Oxygen Lyases - genetics</subject><subject>Carbon-Oxygen Lyases - metabolism</subject><subject>cystathionine γ‐synthase</subject><subject>Feedback, Physiological</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression Regulation, Plant</subject><subject>Metabolism</subject><subject>Metabolism. 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Psychology</topic><topic>Gene Expression Regulation, Plant</topic><topic>Metabolism</topic><topic>Metabolism. Physicochemical requirements</topic><topic>Methionine - metabolism</topic><topic>Methionine - physiology</topic><topic>methionine synthesis</topic><topic>Models, Biological</topic><topic>Molecular Sequence Data</topic><topic>mRNA stability</topic><topic>Nicotiana - genetics</topic><topic>Plant physiology and development</topic><topic>Plants, Genetically Modified - genetics</topic><topic>Plants, Genetically Modified - metabolism</topic><topic>Polyribosomes - metabolism</topic><topic>Protein Structure, Tertiary</topic><topic>RNA Stability</topic><topic>RNA, Messenger - metabolism</topic><topic>Sequence Deletion</topic><topic>sulfur amino acids</topic><topic>transgenic plants</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hacham, Yael</creatorcontrib><creatorcontrib>Schuster, Gadi</creatorcontrib><creatorcontrib>Amir, Rachel</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><jtitle>The Plant journal : for cell and molecular biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hacham, Yael</au><au>Schuster, Gadi</au><au>Amir, Rachel</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An in vivo internal deletion in the N‐terminus region of Arabidopsis cystathionine γ‐synthase results in CGS expression that is insensitive to methionine</atitle><jtitle>The Plant journal : for cell and molecular biology</jtitle><addtitle>Plant J</addtitle><date>2006-03</date><risdate>2006</risdate><volume>45</volume><issue>6</issue><spage>955</spage><epage>967</epage><pages>955-967</pages><issn>0960-7412</issn><eissn>1365-313X</eissn><abstract>Summary Cystathionine γ‐synthase (CGS), the first enzyme of methionine biosynthesis in higher plants, plays an important role in the biosynthesis pathway and in regulating methionine metabolism in plant cells. In response to methionine, the expression of this enzyme is regulated via amino acid sequences located in its N‐terminal. Here, using reverse transcription PCR and ribonuclease protection analysis, we demonstrate that, in addition to the full‐length CGS transcript, a deleted form exists in Arabidopsis. The deleted transcript of CGS that lacks 90 or 87 nt located internally in the regulatory N‐terminal region of CGS maintains the reading frame of the protein. Its association with polyribosomes indicates that this deleted form of CGS is translated. In order to study the function of this deleted form of CGS, we overexpressed it in transgenic tobacco plants. We found that the transgenic plants engineered to express only the deleted form of CGS accumulated methionine to a much higher level than those that expressed the full‐length CGS. Furthermore, in vitro feeding experiments revealed that the deleted form of CGS is not subject to feedback regulation by methionine, as reported for the full‐length transcript. Therefore, although most likely produced from the full‐length CGS, the transcript of the deleted form is insensitive to methionine application and its expression may be important for maintaining methionine metabolism even in the presence of a high level of methionine.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science Ltd</pub><pmid>16507086</pmid><doi>10.1111/j.1365-313X.2006.02661.x</doi><tpages>13</tpages></addata></record>
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subjects Amino Acid Sequence
Arabidopsis
Arabidopsis - enzymology
Arabidopsis - genetics
Base Sequence
Biological and medical sciences
Carbon-Oxygen Lyases - chemistry
Carbon-Oxygen Lyases - genetics
Carbon-Oxygen Lyases - metabolism
cystathionine γ‐synthase
Feedback, Physiological
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation, Plant
Metabolism
Metabolism. Physicochemical requirements
Methionine - metabolism
Methionine - physiology
methionine synthesis
Models, Biological
Molecular Sequence Data
mRNA stability
Nicotiana - genetics
Plant physiology and development
Plants, Genetically Modified - genetics
Plants, Genetically Modified - metabolism
Polyribosomes - metabolism
Protein Structure, Tertiary
RNA Stability
RNA, Messenger - metabolism
Sequence Deletion
sulfur amino acids
transgenic plants
title An in vivo internal deletion in the N‐terminus region of Arabidopsis cystathionine γ‐synthase results in CGS expression that is insensitive to methionine
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