Functional Nonequivalency of Actin Isovariants in<i>Arabidopsis</i>

Kandasamy, Muthugapatti K.; McKinney, Elizabeth C.; Meagher, Richard B.

doi:10.1091/mbc.01-07-0342

Cited by 102 publications

(92 citation statements)

References 39 publications

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“…ACT2 cDNA (At3g18780) was amplified from a previously described plasmid (Kandasamy et al, 2002a) and subcloned as a C-terminal GFP fusion (Chiu et al, 1996) in the plant expression vector. Porin genes At3g01280 (por301) and At5g15090 (hsr2) were amplified from cDNA stocks U12693 and U18357 (Arabidopsis Biological Resource Center), respectively, using the following primers: por301, 5#-GGGATCCCATGGTGAAAGGTCCCGGT and 5#-GAAGGCCTAGGCTTGAGTGCGAGAGCCAATCC; and hsr2, 5#-GTC-AAGGCCTGGGCTTGAGAGCGAGAGCAATC and 5#-GGGATCCATGGT-TAAAGGTCCAGGACTCTACACT.…”

Section: Plasmid and Transgenic Constructsmentioning

confidence: 99%

“…Immunolocalization of HXK1 and F-actin was done using methods previously described (Baskin et al, 1996;Kandasamy et al, 2002a), following leaf cryofixation and acetone freeze substitution of young seedlings. Because anti-HXK1 is a polyclonal antibody and anti-ACT (MAbGPa) is a monoclonal antibody, we labeled cells with corresponding secondary antibodies conjugated to FITC.…”

Section: Cellular Immunolocalizationmentioning

confidence: 99%

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A Role for F-Actin in Hexokinase-Mediated Glucose Signaling

Balasubramanian

Karve²,

Kandasamy³

et al. 2007

Plant Physiology

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HEXOKINASE1 (HXK1) from Arabidopsis (Arabidopsis thaliana) has dual roles in glucose (Glc) signaling and in Glc phosphorylation. The cellular context, though, for HXK1 function in either process is not well understood. Here we have shown that within normal experimental detection limits, AtHXK1 is localized continuously to mitochondria. Two mitochondrial porin proteins were identified as capable of binding to overexpressed HXK1 protein, both in vivo and in vitro. We also found that AtHXK1 can be associated with its structural homolog, F-actin, based on their coimmunoprecipitation from transgenic plants that overexpress HXK1-FLAG or from transient expression assays, and based on their localization in leaf cells after cryofixation. This association might be functionally important because Glc signaling in protoplast transient expression assays is compromised by disruption of F-actin. We also demonstrate that Glc treatment of Arabidopsis seedlings rapidly and reversibly disrupts fine mesh actin filaments. The possible roles of actin in HXK-dependent Glc signaling are discussed.

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Section: Plasmid and Transgenic Constructsmentioning

confidence: 99%

Section: Cellular Immunolocalizationmentioning

confidence: 99%

A Role for F-Actin in Hexokinase-Mediated Glucose Signaling

Balasubramanian

Karve²,

Kandasamy³

et al. 2007

Plant Physiology

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“…Disruption of actin polymerisation by drugs (Baluska et al, 2001), and by some loss-of-function, gain-of-function and misexpression actin mutants (Gilliland et al, 2002;Kandasamy et al, 2002;Nishimura et al, 2003) results in dwarf plants with restricted and uncoordinated cell expansion phenotypes. Sequenced plant genomes contain homologues of many ABPs, some of which have been shown to modulate actin behaviour in planta.…”

Section: Introductionmentioning

confidence: 99%

ArabidopsisCAP1 – a key regulator of actin organisation and development

Deeks

Rodrigues

Dimmock

et al. 2007

Journal of Cell Science

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Maintenance of F-actin turnover is essential for plant cell morphogenesis. Actin-binding protein mutants reveal that plants place emphasis on particular aspects of actin biochemistry distinct from animals and fungi. Here we show that mutants in CAP1, an A. thaliana member of the cyclase-associated protein family, display a phenotype that establishes CAP1 as a fundamental facilitator of actin dynamics over a wide range of plant tissues. Plants homozygous for cap1 alleles show a reduction in stature and morphogenetic disruption of multiple cell types. Pollen grains exhibit reduced germination efficiency, and cap1 pollen tubes and root hairs grow at a decreased rate and to a reduced length. Live cell imaging of growing root hairs reveals actin filament disruption and cytoplasmic disorganisation in the tip growth zone. Mutant cap1 alleles also show synthetic phenotypes when combined with mutants of the Arp2/3 complex pathway, which further suggests a contribution of CAP1 to in planta actin dynamics. In yeast, CAP interacts with adenylate cyclase in a Ras signalling cascade; but plants do not have Ras. Surprisingly, cap1 plants show disruption in plant signalling pathways required for co-ordinated organ expansion suggesting that plant CAP has evolved to attain plant-specific signalling functions.

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“…Ectopic overexpression of fl-cDNAs often causes their misexpression, and the altered phenotypes appearing in the FOX plants are not necessarily related to the original function of the transgene (Kandasamy et al 2002, Newman et al 2004. To better understand the native function of a gene, it is sometimes necessary to obtain different types of mutants or transgenic lines in addition to the FOX lines and to understand the expression profiles of genes of interest (Sato et al 2010).…”

Section: Rice Fox-rice Resourcesmentioning

confidence: 99%

Rice transgenic resources with gain-of-function phenotypes

et al. 2010

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After the completion of its genomic sequencing, rice (Oryza sativa L.) has become firmly established as the premiere model plant among monocot crops. Various genetic resources have been developed for rice to accelerate the identification and characterization of rice genes with as-yet unknown functions. These resources include collections that carry natural variations and progeny derived from chemical and irradiation mutagenesis. Collections of rice strains with natural or artificial DNA insertions that create loss-of-function mutations have been of great importance for gene discovery and tagging. However, it is often difficult to observe mutant phenotypes for functionally duplicated genes and genes essential for plant growth and development. To overcome this disadvantage, gain-of-function strategies have been developed, including activation tagging and full-length cDNA overexpressing gene (FOX)-hunting systems. In this review, we summarize the current status and discuss the advantages and disadvantages of the gain-of-function approaches, and then, perspectives on accelerating discovery of rice genes.

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Functional Nonequivalency of Actin Isovariants inArabidopsis

Cited by 102 publications

References 39 publications

A Role for F-Actin in Hexokinase-Mediated Glucose Signaling

A Role for F-Actin in Hexokinase-Mediated Glucose Signaling

ArabidopsisCAP1 – a key regulator of actin organisation and development

Rice transgenic resources with gain-of-function phenotypes

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