IMPa-4, an<i>Arabidopsis</i>Importin α Isoform, Is Preferentially Involved in<i>Agrobacterium</i>-Mediated Plant Transformation

Bhattacharjee, Saikat; Lee, Lan‐Ying; Oltmanns, Heiko; Cao, Hongbin; Veena,; Cuperus, Josh T.; Gelvin, Stanton B.

doi:10.1105/tpc.108.060467

Cited by 112 publications

(187 citation statements)

References 89 publications

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“…5B). Interestingly, this does not correlate with the phylogenetic relation between importin ␣ proteins (46).…”

Section: N-terminal Region Of Atrem13 Is Intrinsically Disordered-mentioning

confidence: 90%

“…Importin ␣ proteins are soluble receptors involved in nuclear trafficking. In A. thaliana, this family comprises 9 members, some of which are involved in interaction with VirD2 and VirE2 bacterial T-complex proteins and mediate nuclear localization of these bacterial proteins (46). Moreover, AtIMPa3 (MOS6) plays a role in plant innate immunity because it is a suppressor of resistance responses and immunity to pathogens (42), further supporting possible roles of remorin proteins during plant-microbe interactions and innate immunity.…”

Section: Discussionmentioning

confidence: 99%

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The Intrinsically Disordered N-terminal Region of AtREM1.3 Remorin Protein Mediates Protein-Protein Interactions

Marín

Thallmair

Ott

2012

Journal of Biological Chemistry

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“…5B). Interestingly, this does not correlate with the phylogenetic relation between importin ␣ proteins (46).…”

Section: N-terminal Region Of Atrem13 Is Intrinsically Disordered-mentioning

confidence: 90%

Section: Discussionmentioning

confidence: 99%

The Intrinsically Disordered N-terminal Region of AtREM1.3 Remorin Protein Mediates Protein-Protein Interactions

Marín

Thallmair

Ott

2012

Journal of Biological Chemistry

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“…To gain better understanding on the functional significance of the VirE2 interaction with XRCC4 in relation to Agrobacteriummediated transformation, we took advantage of transgenic Arabidopsis plants expressing VirE2-YFP (Bhattacharjee et al, 2008). We hypothesized that VirE2 can interact with XRCC4 in plants expressing VirE2-YFP, thereby delaying the NHEJ-mediated DSB repair and providing greater opportunity for T-DNA integration.…”

Section: Arabidopsis Plants Expressing Vire2 Are Hypersensitive To Blmentioning

confidence: 99%

“…Many molecules of Agrobacterium VirE2 protein likely enter the plant cell independent of T-DNA (Sundberg et al, 1996;Gelvin, 1998) and may localize into the nucleus (Citovsky et al, 1992). VirE2 localization in plant cells, either to the nucleus or perinuclear vicinity, is controversial (Citovsky et al, 1992;Bhattacharjee et al, 2008). We propose that free VirE2 molecules can interact with XRCC4 and titrate/exclude active XRCC4 protein available for DSB repair in the cell.…”

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confidence: 99%

Agrobacterium May Delay Plant Nonhomologous End-Joining DNA Repair via XRCC4 to Favor T-DNA Integration

et al. 2012

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Agrobacterium tumefaciens is a soilborne pathogen that causes crown gall disease in many dicotyledonous plants by transfer of a portion of its tumor-inducing plasmid (T-DNA) into the plant genome. Several plant factors that play a role in Agrobacterium attachment to plant cells and transport of T-DNA to the nucleus have been identified, but the T-DNA integration step during transformation is poorly understood and has been proposed to occur via nonhomologous end-joining (NHEJ)-mediated doublestrand DNA break (DSB) repair. Here, we report a negative role of X-RAY CROSS COMPLEMENTATION GROUP4 (XRCC4), one of the key proteins required for NHEJ, in Agrobacterium T-DNA integration. Downregulation of XRCC4 in Arabidopsis and Nicotiana benthamiana increased stable transformation due to increased T-DNA integration. Overexpression of XRCC4 in Arabidopsis decreased stable transformation due to decreased T-DNA integration. Interestingly, XRCC4 directly interacted with Agrobacterium protein VirE2 in a yeast two-hybrid system and in planta. VirE2-expressing Arabidopsis plants were more susceptible to the DNA damaging chemical bleomycin and showed increased stable transformation. We hypothesize that VirE2 titrates or excludes active XRCC4 protein available for DSB repair, thus delaying the closure of DSBs in the chromosome, providing greater opportunity for T-DNA to integrate.

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“…Following transfer of T-DNA to the plant cytoplasm, it likely forms complexes with various transferred Agrobacterium virulence effector proteins, including VirE2, a single-stranded DNA binding protein (Gietl et al, 1987;Christie et al, 1988;Citovsky et al, 1988Citovsky et al, , 1992Das, 1988) and VirF, which may mediate proteolysis of VirE2 in the nucleus (Schrammeijer et al, 2001;Tzfira et al, 2004). In addition, plantencoded proteins, such as VIP1 and importin a, may form supercomplexes with the T-strand and help target T-DNA to the nucleus (Ballas and Citovsky, 1997;Tzfira et al, 2001;Bhattacharjee et al, 2008;Lee et al, 2008). The timing and subcellular location of VirE2 interaction with T-strands remains unknown.…”

Section: Overexpression Of a Hta1 Cdna Can Increase Expression Of A Smentioning

confidence: 99%

Overexpression of SeveralArabidopsisHistone Genes IncreasesAgrobacterium-Mediated Transformation and Transgene Expression in Plants

et al. 2009

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The Arabidopsis thaliana histone H2A-1 is important for Agrobacterium tumefaciens-mediated plant transformation. Mutation of HTA1, the gene encoding histone H2A-1, results in decreased T-DNA integration into the genome of Arabidopsis roots, whereas overexpression of HTA1 increases transformation frequency. To understand the mechanism by which HTA1 enhances transformation, we investigated the effects of overexpression of numerous Arabidopsis histones on transformation and transgene expression. Transgenic Arabidopsis containing cDNAs encoding histone H2A (HTA), histone H4 (HFO), and histone H3-11 (HTR11) displayed increased transformation susceptibility, whereas histone H2B (HTB) and most histone H3 (HTR) cDNAs did not increase transformation. A parallel increase in transient gene expression was observed when histone HTA, HFO, or HTR11 overexpression constructs were cotransfected with double-or single-stranded forms of a gusA gene into tobacco (Nicotiana tabacum) protoplasts. However, these cDNAs did not increase expression of a previously integrated transgene. We identified the N-terminal 39 amino acids of H2A-1 as sufficient to increase transient transgene expression in plants. After transfection, transgene DNA accumulates more rapidly in the presence of HTA1 than with a control construction. Our results suggest that certain histones enhance transgene expression, protect incoming transgene DNA during the initial stages of transformation, and subsequently increase the efficiency of Agrobacterium-mediated transformation. INTRODUCTIONAgrobacterium tumefaciens-mediated plant genetic transformation is an important experimental tool for investigation of various aspects of plant biology and for agricultural biotechnology. Development of this transformation technology represents the culmination of many decades of effort to improve tissue culture and plant genetic engineering techniques. Agrobacteriummediated transformation is based on a conjugative transfer-like process, which eventually takes the T-DNA to the host cell nucleus (Gelvin, 2000(Gelvin, , 2003a(Gelvin, , 2009Tzfira and Citovsky, 2003;Citovsky et al., 2007). After attachment of the bacterium to plant cells and induction of Agrobacterium virulence (vir) genes, a single-stranded DNA (the T-strand) is processed from the resident tumor-inducing or root-inducing plasmid and transported from the bacterium to the plant cell. In addition, a number of Virulence effector proteins, including VirD2 (attached to the T-strand), the single-strand DNA binding protein VirE2, plus VirE3, VirD5, and VirF are also transferred to the plant (Otten et al., 1984;Stahl et al., 1998;Vergunst et al., 2000Vergunst et al., , 2003Vergunst et al., , 2005Schrammeijer et al., 2003). These proteins are likely involved in protecting T-DNA from nuclease digestion, directing T-DNA to the nucleus, stripping proteins from the T-strand prior to integration, and integrating T-DNA into the plant genome. T-DNA integration occurs randomly into genomic DNA by the process of illegitimate recombination (...

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IMPa-4, anArabidopsisImportin α Isoform, Is Preferentially Involved inAgrobacterium-Mediated Plant Transformation

Cited by 112 publications

References 89 publications

The Intrinsically Disordered N-terminal Region of AtREM1.3 Remorin Protein Mediates Protein-Protein Interactions

The Intrinsically Disordered N-terminal Region of AtREM1.3 Remorin Protein Mediates Protein-Protein Interactions

Agrobacterium May Delay Plant Nonhomologous End-Joining DNA Repair via XRCC4 to Favor T-DNA Integration

Overexpression of SeveralArabidopsisHistone Genes IncreasesAgrobacterium-Mediated Transformation and Transgene Expression in Plants

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