Genomic organization and biosynthesis of secreted and cytoplasmic forms of gelsolin

Kwiatkowski, David J.; Mehl, Ryan A.; Hl, Yin

doi:10.1083/jcb.106.2.375

Cited by 151 publications

(88 citation statements)

References 41 publications

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“…We first carried out direct sequencing of the gelsolin promoter (from Ϫ580 to 75, relative to the transcription start site) using the PCR products derived from human bladder cancer cells (DAB-1, KU-7 and UMUC-2) and the cells derived from normal uroepithelium (SV-HUC-1). Although it was found that the gelsolin promoter of cancer and normal tissue-derived cells were identical in sequence, this sequence differed from that previously reported at 2 nucleotide positions: Ϫ153 G (from K, which shows G or T in the original report 24 ) and Ϫ170 T (our addition) from the transcription start site (Fig. 1).…”

Section: Mutation and Methylationcontrasting

confidence: 76%

Gelsolin gene silencing involving unusual hypersensitivities to dimethylsulfate and KMnO₄in vivo footprinting on its promoter region

Haga

Fujita

Nomoto

et al. 2004

Intl Journal of Cancer

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We previously reported that gelsolin gene expression is reduced in various tumors. In an effort to gain further insights into the mechanism of gelsolin downregulation in tumors, we examined the in vivo properties of the gelsolin promoter in urinary bladder cancer cell lines. Neither mutation nor hypermethylation was responsible for gene silencing at the promoter. After exposure to trichostatin A (TSA), a histone deacetylase inhibitor, gelsolin promoter activity was markedly enhanced in the cancer cells, not in cells derived from normal tissue. Chromatin immunoprecipitation assays revealed that both histones H3 and H4 were hypoacetylated in the promoter region of the cancer cells, and the accumulation of acetylated histones was detected by TSA treatment. Key words: gelsolin; gene silencing; dimethylsulfate hypersensitivity; bladder cancer; in vivo footprintingCancers occur due to the accumulation of abnormalities in gene function. While many of them are genetic (mutation and deletion), epigenetically mediated changes in gene expression have been increasingly investigated. Hypermethylation of CpG islands, which are GC-rich regions usually located at the 5Ј end of genes, results in alterations in the histone acetylation and methylation status around gene promoter regions. Aberrant cytosine methylation and concomitant histone modifications play important roles in the repression of over half of the tumor suppressor genes during malignant transformation. 1,2Previously, we reported that the expression of gelsolin, an actin-regulatory protein, is frequently silenced in various cancers, including stomach, colon, urinary bladder and lung, and that gelsolin functions as a tumor suppressor. [3][4][5][6][7] These findings were also demonstrated in breast and prostate cancers. 8,9 On the other hand, a subset of non small cell lung cancers is characterized by high expression levels of gelsolin correlated with lymphatic invasion, 10 and there was a gradual increase in gelsolin with an increase in tumor grade and stage in uroepithelial carcinoma. 11 Gelsolin induced invasion of epithelial cells as well as colon adenocarcinoma cells. 12 It is therefore possible that a decrease in expression during a particular stage is followed by an increase during another stage. To understand roles of gelsolin in the whole process of tumorigenesis, mechanisms for both down-and upregulations should be analyzed. In breast cancers, gelsolin genes were not mutated; however, epigenetic changes in the chromatin structure were responsible for the deficiency. 13 Histone deacetylase (HDAC) inhibitors, such as trichostatin A (TSA) and apicidin, selectively induced the expression of gelsolin. 14,15 Recent studies have suggested a new mechanism for cell-specific gene regulation linking nuclear architecture, chromatin structure and functional organization of DNA sequences. 16 In an effort to determine the molecular basis of gelsolin downregulation in urinary bladder cancer and to better understand the carcinogenic process, we investigated the gelsolin prom...

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Section: Mutation and Methylationcontrasting

confidence: 76%

Gelsolin gene silencing involving unusual hypersensitivities to dimethylsulfate and KMnO₄in vivo footprinting on its promoter region

Haga

Fujita

Nomoto

et al. 2004

Intl Journal of Cancer

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“…Gelsolin segment 5 (amino acid sequence 516-618), segment 5-6 (amino acid sequence 516-731) and segment 6 (amino acid sequence 640-731) [15,16,29] were obtained by PCR amplification with LKCG as a template. Each PCR product was subcloned into the LK444 expression plasmid (a kind gift from P. Gunning) [13,20] after insertion of a FLAG tag sequence at the 5' termini or into EcoRI-XhoI site of pCMV-FLAG expression plasmid which consists of pcDNA3.1(+) Invitrogen, Carlsbad, CA and 3xFLAG tag.…”

Section: Plasmids Cells and Transfectionmentioning

confidence: 99%

“…Gelsolin, an actin-regulatory protein that modulates actin assembly and disassembly, is found as both an intrinsic cytoplasmic protein and as a secreted plasma protein [15,16]. Immunocytochemical studies using anti-gelsolin antibody revealed that the protein is localized in filopodia and the body part of growth cones of neuronally differentiated rat pheochromocytoma (PC12) cells treated with nerve growth factor (NGF) [31].…”

Section: Introductionmentioning

confidence: 99%

Inhibition of Alzheimer's amyloid-β peptide-induced reduction of mitochondrial membrane potential and neurotoxicity by gelsolin

Qiao¹,

Koya²,

Nakagawa³

et al. 2005

Neurobiology of Aging

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Amyloid-beta (Abeta) peptides play a central role in the development of Alzheimer's disease. They are known to induce mitochondrial dysfunction and caspase activation, resulting in apoptosis of neuronal cells. Here we show that human cytoplasmic gelsolin inhibits Abeta peptide-induced cell death of neuronally differentiated rat pheochromocytoma (PC-12) cells. We also show that the segment 5 but not 6 of human cytoplasmic gelsolin is the important region responsible for inhibition of Abeta-induced cytotoxicity. Mitochondrial dysfunction associated with cell death, membrane potential loss and the release of cytochrome c are all abrogated in the presence of human full-length or segment 5 cytoplasmic gelsolin. Furthermore, RNA interference to reduce expression of endogenous gelsolin in PC12 cells shows that rat gelsolin act as an inhibitor of Abeta cytotoxicity. These results demonstrate that cytoplasmic gelsolin plays a important role in inhibiting Abeta-induced cytotoxicity by inhibiting apoptotic mitochondrial changes. The segment 5 of human cytoplasmic gelsolin is sufficient for the function.

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“…This is comparable to the amount of the 40-kDa earthworm actin modulator in annelid muscle [13], but significantly higher than that calculated for mammalian skeletal muscle [44, 4.51. Despite its low content in cytoplasmic gelsolin [46], mammalian muscle synthesizes relatively large amounts of the slightly different plasma gelsolin variant [24,47,48]. We assume that the purified crayfish modulator is exclusively cytoplasmic, since the protein appeared as a single component in two-dimensional electrophoresis.…”

Section: Discussionmentioning

confidence: 99%

“…Sequence analysis revealed a characteristic sixfold repeated pattern in the gelsolin-type proteins [15, 18, 211 and a threefold repeat in severin and fragmin with high similarity to one half of the gelsolin molecule [17], thus indicating that the gelsolin-like proteins may have evolved from severin-like proteins by gene duplication [16,22,231. It was initially assumed that the 80-ma-type proteins exclusively occur in vertebrates [24]. However, recent findings of a 110-kDa villin-like protein in sea urchin eggs [25] and a gelsolin-like protein in Drosophila [26] suggest a more complex situation where both types of protein may occur within the same phylum.…”

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confidence: 99%

A gelsolin‐related actin‐severing protein with fully reversible actin‐binding properties from the tail muscle of crayfish, Astacus leptodactylus

Bock

Hinssen

D’Haese

1994

European Journal of Biochemistry

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A Ca2+-dependent actin-severing protein was purified from the tail muscle of the crayfish Astacus leptodactylus. The isolation procedure involved extraction at low ionic strength in the presence of EGTA, followed by ammonium sulfate fractionation, ion-exchange chromatography and gel filtration. The purified crayfish actin modulator appeared as a single band with a molecular mass of 105 kDa on SDSPAGE. The crustacean actin modulator revealed basic functional properties in common with vertebrate gelsolin, like the Ca"-activated severing of F-actin and the nucleation of actin polymerization. However, both proteins differed in major aspects: Caz+ activation of crayfish actin modulator started at lower threshold concentrations (0.1 pM). The effect of the modulator on shortening the nucleation phase of actin polymerization was significantly weaker at lower modulatorhctin ratios. The modulator formed three distinct stoichiometric complexes with G-actin, identified as binary, ternary and quaternary. Binding of G-actin occurred in a low cooperative manner and was completely reversible by EGTA. Despite some properties being similar to those of villin, crayfish actin modulator did not cross-link actin filaments. It is regarded in principle as a gelsolintype protein, but with characteristic functional deviations from vertebrate gelsolin.A large number of actin-associated proteins mediate the stability and dynamics of the cytoskeletal architecture as well as the generation of motive force in both muscle and nonmuscle cells (for reviews see . A distinct class of actin-binding proteins, initially termed actin modulators [ 5 ] , comprises proteins of very different evolutionary origin like gelsolin in vertebrates [6-91, severin [lo] and fragmin [5,11, 121 in slime moulds or earthworm actin modulator [13] in annelids. Nevertheless, all these proteins are remarkably similar both in structure and in function [ 13 -181. Their characteristic pattern of interaction with actin is the Ca2+-dependent severing of actin filaments, the promotion of nucleation of actin polymerization and a capping of the rapidly polymerizing end of F-actin.Apart from these properties, distinct differences exist between actin-modulating proteins. First, they vary in molecular mass over 85-90 kDa for the gelsolidvillin type [7,19, 201 to 40 kDa for severin, fragmin and the corresponding protein from annelid muscle. Sequence analysis revealed a characteristic sixfold repeated pattern in the gelsolin-type proteins [15, 18, 211 and a threefold repeat in severin and fragmin with high similarity to one half of the gelsolin molecule [17], thus indicating that the gelsolin-like proteins may have evolved from severin-like proteins by gene duplication [16, 22, 231.

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Genomic organization and biosynthesis of secreted and cytoplasmic forms of gelsolin

Cited by 151 publications

References 41 publications

Gelsolin gene silencing involving unusual hypersensitivities to dimethylsulfate and KMnO₄in vivo footprinting on its promoter region

Gelsolin gene silencing involving unusual hypersensitivities to dimethylsulfate and KMnO₄in vivo footprinting on its promoter region

Inhibition of Alzheimer's amyloid-β peptide-induced reduction of mitochondrial membrane potential and neurotoxicity by gelsolin

A gelsolin‐related actin‐severing protein with fully reversible actin‐binding properties from the tail muscle of crayfish, Astacus leptodactylus

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Genomic organization and biosynthesis of secreted and cytoplasmic forms of gelsolin

Cited by 151 publications

References 41 publications

Gelsolin gene silencing involving unusual hypersensitivities to dimethylsulfate and KMnO4in vivo footprinting on its promoter region

Gelsolin gene silencing involving unusual hypersensitivities to dimethylsulfate and KMnO4in vivo footprinting on its promoter region

Inhibition of Alzheimer's amyloid-β peptide-induced reduction of mitochondrial membrane potential and neurotoxicity by gelsolin

A gelsolin‐related actin‐severing protein with fully reversible actin‐binding properties from the tail muscle of crayfish, Astacus leptodactylus

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Gelsolin gene silencing involving unusual hypersensitivities to dimethylsulfate and KMnO₄in vivo footprinting on its promoter region

Gelsolin gene silencing involving unusual hypersensitivities to dimethylsulfate and KMnO₄in vivo footprinting on its promoter region