Serveur d'exploration sur l'agrobacterium et la transgénèse

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Arabidopsis VIRE2 INTERACTING PROTEIN2 is required for Agrobacterium T-DNA integration in plants.

Identifieur interne : 000726 ( Main/Exploration ); précédent : 000725; suivant : 000727

Arabidopsis VIRE2 INTERACTING PROTEIN2 is required for Agrobacterium T-DNA integration in plants.

Auteurs : Ajith Anand [États-Unis] ; Alexander Krichevsky ; Sebastian Schornack ; Thomas Lahaye ; Tzvi Tzfira ; Yuhong Tang ; Vitaly Citovsky ; Kirankumar S. Mysore

Source :

RBID : pubmed:17496122

Descripteurs français

English descriptors

Abstract

Agrobacterium tumefaciens-mediated genetic transformation is an efficient tool for genetic engineering of plants. VirE2 is a single-stranded DNA binding Agrobacterium protein that is transported into the plant cell and presumably protects the T-DNA from degradation. Using a yeast two-hybrid system, we identified Arabidopsis thaliana VIRE2-INTERACTING PROTEIN2 (VIP2) with a NOT domain that is conserved in both plants and animals. Furthermore, we provide evidence supporting VIP2 interaction with VIP1, a basic domain/leucine zipper motif-containing protein required for nuclear import and integration of T-DNA. Virus-induced gene silencing of VIP2 in Nicotiana benthamiana and characterization of the Arabidopsis vip2 mutant (At vip2) demonstrate that VIP2 is required for Agrobacterium-mediated stable transformation but not for transient transformation. Assays based upon a promoter-trap vector and quantification of T-DNA integration further confirmed VIP2 involvement in T-DNA integration. Interestingly, VIP2 transcripts were induced to a greater extent over prolonged periods after infection with a T-DNA transfer-competent Agrobacterium strain compared with the transfer-deficient Agrobacterium strain. Transcriptome analyses of At vip2 suggest that VIP2 is likely a transcriptional regulator, and the recalcitrancy to transformation in At vip2 is probably due to the combination of muted gene expression response upon Agrobacterium infection and repression of histone genes resulting in decreased T-DNA integration events.

DOI: 10.1105/tpc.106.042903
PubMed: 17496122
PubMed Central: PMC1913729


Affiliations:


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Le document en format XML

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<term>Agrobacterium tumefaciens (metabolism)</term>
<term>Amino Acid Sequence (MeSH)</term>
<term>Arabidopsis (genetics)</term>
<term>Arabidopsis (metabolism)</term>
<term>Arabidopsis (microbiology)</term>
<term>Arabidopsis Proteins (chemistry)</term>
<term>Arabidopsis Proteins (genetics)</term>
<term>Arabidopsis Proteins (metabolism)</term>
<term>Bacterial Proteins (metabolism)</term>
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<term>DNA, Bacterial (metabolism)</term>
<term>DNA-Binding Proteins (metabolism)</term>
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<term>Gene Silencing (MeSH)</term>
<term>Histones (genetics)</term>
<term>Ion Channels (metabolism)</term>
<term>Molecular Sequence Data (MeSH)</term>
<term>Mutation (genetics)</term>
<term>Oligonucleotide Array Sequence Analysis (MeSH)</term>
<term>Protein Binding (MeSH)</term>
<term>RNA, Messenger (genetics)</term>
<term>RNA, Messenger (metabolism)</term>
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<term>Tobacco (metabolism)</term>
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<term>Agrobacterium tumefaciens (métabolisme)</term>
<term>Analyse de profil d'expression de gènes (MeSH)</term>
<term>Arabidopsis (génétique)</term>
<term>Arabidopsis (microbiologie)</term>
<term>Arabidopsis (métabolisme)</term>
<term>Canaux ioniques (métabolisme)</term>
<term>Données de séquences moléculaires (MeSH)</term>
<term>Extinction de l'expression des gènes (MeSH)</term>
<term>Facteurs généraux de transcription (MeSH)</term>
<term>Histone (génétique)</term>
<term>Liaison aux protéines (MeSH)</term>
<term>Mutation (génétique)</term>
<term>Noyau de la cellule (métabolisme)</term>
<term>Protéines bactériennes (métabolisme)</term>
<term>Protéines d'Arabidopsis (composition chimique)</term>
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<term>Protéines d'Arabidopsis (métabolisme)</term>
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<term>Protéines de transport (métabolisme)</term>
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<term>Régulation de l'expression des gènes végétaux (MeSH)</term>
<term>Séquence d'acides aminés (MeSH)</term>
<term>Séquençage par oligonucléotides en batterie (MeSH)</term>
<term>Tabac (microbiologie)</term>
<term>Tabac (métabolisme)</term>
<term>Transcription génétique (MeSH)</term>
<term>Transformation génétique (MeSH)</term>
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<term>Arabidopsis Proteins</term>
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<term>Protéines d'Arabidopsis</term>
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<term>Arabidopsis</term>
<term>Arabidopsis Proteins</term>
<term>Bacterial Proteins</term>
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<term>Cell Nucleus</term>
<term>DNA, Bacterial</term>
<term>DNA-Binding Proteins</term>
<term>Ion Channels</term>
<term>RNA, Messenger</term>
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<term>Tabac</term>
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<term>ARN messager</term>
<term>Agrobacterium tumefaciens</term>
<term>Arabidopsis</term>
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<term>Noyau de la cellule</term>
<term>Protéines bactériennes</term>
<term>Protéines d'Arabidopsis</term>
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<term>Amino Acid Sequence</term>
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<term>Oligonucleotide Array Sequence Analysis</term>
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<term>Reproducibility of Results</term>
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<term>Reproductibilité des résultats</term>
<term>Régulation de l'expression des gènes végétaux</term>
<term>Séquence d'acides aminés</term>
<term>Séquençage par oligonucléotides en batterie</term>
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<div type="abstract" xml:lang="en">Agrobacterium tumefaciens-mediated genetic transformation is an efficient tool for genetic engineering of plants. VirE2 is a single-stranded DNA binding Agrobacterium protein that is transported into the plant cell and presumably protects the T-DNA from degradation. Using a yeast two-hybrid system, we identified Arabidopsis thaliana VIRE2-INTERACTING PROTEIN2 (VIP2) with a NOT domain that is conserved in both plants and animals. Furthermore, we provide evidence supporting VIP2 interaction with VIP1, a basic domain/leucine zipper motif-containing protein required for nuclear import and integration of T-DNA. Virus-induced gene silencing of VIP2 in Nicotiana benthamiana and characterization of the Arabidopsis vip2 mutant (At vip2) demonstrate that VIP2 is required for Agrobacterium-mediated stable transformation but not for transient transformation. Assays based upon a promoter-trap vector and quantification of T-DNA integration further confirmed VIP2 involvement in T-DNA integration. Interestingly, VIP2 transcripts were induced to a greater extent over prolonged periods after infection with a T-DNA transfer-competent Agrobacterium strain compared with the transfer-deficient Agrobacterium strain. Transcriptome analyses of At vip2 suggest that VIP2 is likely a transcriptional regulator, and the recalcitrancy to transformation in At vip2 is probably due to the combination of muted gene expression response upon Agrobacterium infection and repression of histone genes resulting in decreased T-DNA integration events.</div>
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