title: A cotton LIM domain-containing protein (GhWLIM5) is involved in bundling actin filaments.
abstract: LIM-domain proteins play important roles in cellular processes in eukaryotes. In this study, a LIM protein gene, GhWLIM5, was identified in cotton. Quantitative RT-PCR analysis showed that GhWLIM5 was expressed widely in different cotton tissues and had a peak in expression during fiber elongation. GFP fluorescence assay revealed that cotton cells expressing GhWLIM5:eGFP fusion gene displayed a network distribution of eGFP fluorescence, suggesting that GhWLIM5 protein is mainly localized to the cell cytoskeleton. When GhWLIM5:eGFP transformed cells were stained with rhodamine-phalloidin there was consistent overlap in eGFP and rhodamine-palloidin signals, demonstrating that GhWLIM5 protein is colocalized with the F-actin cytoskeleton. In addition, high-speed cosedimentation assay verified that GhWLIM5 directly bound actin filaments, while low cosedimentation assay and microscopic observation indicated that GhWLIM5 bundled F-actin in vitro. Increasing amounts of GhWLIM5 protein were able to protect F-actin from depolymerization in vitro in the presence of Lat B (an F-actin depolymerizer). Our results contribute to a better understanding of the biochemical role of GhWLIM5 in modulating the dynamic F-actin network in cotton.
authors: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Sciences, Central China Normal University, Wuhan 430079, China.
authors: Yang
authors: Y
authors: Li
authors: Jia
authors: J
authors: Jiang
authors: Lan
authors: L
authors: Li
authors: Xiu-Lan
authors: XL
authors: Wang
authors: Na-Na
authors: NN
authors: Wang
authors: Deng-Di
authors: DD
authors: Li
authors: Xue-Bao
authors: XB
authors: Li
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000199:Actins / Q000378:metabolism*; D000595:Amino Acid Sequence; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D003368:Gossypium / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D060588:LIM Domain Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010590:Phalloidine / Q000031:analogs & derivatives / Q000378:metabolism; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012235:Rhodamines / Q000378:metabolism
chemical_list: D000199:Actins; D060588:LIM Domain Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D012235:Rhodamines; C485184:enhanced green fluorescent protein; C504731:rhodamine-phalloidin; D049452:Green Fluorescent Proteins; D010590:Phalloidine