• Molecules and Cells
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• v.21 no.1
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• pp.76-81
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• 2006

Tubulin is synthesized in the cell body and must be delivered to the axon to support axonal growth. However, the exact form in which these proteins, in particular tubulin, move within the axon remains contentious. According to the "polymer transport model", tubulin is transported in the form of microtubules. In an alternative hypothesis, the "short oligomer transport model", tubulin is added to existing, stationary microtubules along the axon. In this study, we measured the translocation of microtubule plus ends in soma segments, the middle of axonal shafts and the growth cone areas, by expressing GFP-EB3 in cultured Xenopus embryonic spinal neurons. We found that none of the microtubules in the three compartments were transported rapidly as would be expected from the polymer transport model. These results suggest that microtubules are stationary in most segments of the axon, thus supporting the model according to which tubulin is transported in nonpolymeric form in rapidly growing Xenopus neurons.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.1
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• pp.361-365
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• 2012

Aims: To investigate the relationship between the expression of ${\beta}$-tubulin III and survivin in advanced breast cancers and chemotherapeutic effects of docetaxel. Methods: Clinical pathological data of 74 patients with advanced breast cancer were retrospectively analyzed after docetaxel chemotherapy. Expression of ${\beta}$-tubulin III and survivin was assessed by immunohistochemistry and analyzed with reference to therapeutical and adverse effects of docetaxel. Results: The positive expression rate of ${\beta}$-tubulin III was 38.1% (32/84), while that of survivin was 76.2% (64/84). The effective rate (complete response + partial response) was 52.4%. That for patients with the positive expression of ${\beta}$-tubulin III or/and survivin was significantly lower than for those with negative expression (P<0.05). There were significant differences in the non-progression of median diseases, 1-year and 2-year survival rates of between the patients with positive and negative expression (P<0.05). The main side effects were myelosuppression, alimentary canal response and alopecie, no differences being observed between groups. Conclusions: The combined detection of ${\beta}$-tubulin III and survivin is a predictive index for chemotherapy effects of docetaxel in metastatic breast cancer.

• International Journal of Industrial Entomology and Biomaterials
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• v.11 no.1
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• pp.37-42
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• 2005

We cloned the $\beta$-tubulin genes from the wild type strain and two benomyl-resistant mutants of Metahizium anisopliae and determined their nucleotide sequences. A $\beta$-tubulin encoding 448-residue protein from wild type M. anisopliae shows strong homology to other $\beta$-tubulins. The coding region is interrupted by four introns. Comparisons of intron position between the M. anisopliae gene and other fungal $\beta$-tubulin genes show considerable positional conservation. The mutations responsible for benomyl resistance were determined in two spontaneous mutants, 8-18 and 8­19. One mutant 8-18 substituted glutamate for aspar­agine at position 33 and lysine for glutamine at position 134. The other mutant 8-19 showed alterations at three positions of $\beta$-tubulin arginine for tryptophan at position 21, lysine for asparagine at position 33, and phenylalanine for leucine at position 240. These data suggest that regions of $\beta$-tubulin containing amino acids 21, 33,134, and 240 interact to form the binding site of benomyl.

• BMB Reports
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• v.41 no.1
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• pp.62-67
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• 2008

The present study was aimed to elucidate the mechanism of stabilization of tubulin by deuterium oxide ($D_2O$). Rate of decrease of tryptophan fluorescence during aging of tubulin at 4$^{\circ}C$ and 37$^{\circ}C$ was significantly lower in $D_2O$ than in $H_2O$. Circular dichroism spectra of tubulin after incubation at 4$^{\circ}C$, suggested that complete stabilization of the secondary structure in D2O during the first 24 hours of incubation. The number of available cysteine measured by DTNB reaction was decreased to a lesser extent in $D_2O$ than in $H_2O$. . During the increase in temperature of tubulin, the rate of decrease of fluorescence at 335 nm and change of CD value at 222 nm was lesser in $D_2O$. Differential Scanning calorimetric experiments showed that the $T_m$ values for tubulin unfolding in $D_2O$ were 58.6$^{\circ}C$ and 62.17$^{\circ}C$, and in $H_2O$. those values were 55.4$^{\circ}C$ and 59.35$^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• v.12 no.4
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• pp.628-634
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• 2002

A microfilament-based motility in Toxoplasma gondii (T. gondii) Is involved in host cell invasion, yet the exact mechanism has not yet been determined. Accordingly, the current study examined the localization of actin and tubulin in T gondii using immunofluorescent (IF) and immunogold staining for electron microscopy. Indirect immunofluorescence (IF) staining using anti-actin and anti-tubulin monoclonal antibodies (mAbs) revealed localization of fluorescence on the entire surface of the tachyzoites. The actin in T. gondii was observed by immunogold staining, and the gold particles were seen on the surface, especially at the anterior end and in the cytoplasm of the parasite. However, there were no gold particles in the nucleus, rhoptries, and dense granules. The tubulin in T gondii was located on the surface and in the cytoplasm of the tachyzoites in the extracellular parasite, compared with anterior part of tachyzoites in the intracellular parasite. The antigens of T gondii recognized by anti-actin mAb were 107 kDa, 50 kDa, 48 kDa, and 40 kDa proteins, while those recognized by anti-tubulin mAb were 56 kDa, 52 kDa, and 34 kDa proteins. Tachyzoites of T gondii pretreated with the actin inhibitor, cytochalasin D (20 $\mu\textrm{g}$/ml), and tubulin inhibitor, colchicine (2$\times$10$\^$-6/ M), for 30 min at 37$\^{C}$ were used to infect the isolated mouse macrophages (tachyzo ites:macrophage=2:1). Pretreatment with the inhibitors resulted in lower multiplication of tachyzoites within the macrophages than in the untreated group 18 h post infection (p<0.05). Therefore, the present results suggest that actin and tubulin appear to be involved in the invasion of and multiplication in host cells.

• Applied Microscopy
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• v.28 no.2
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• pp.193-205
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• 1998

The Urechis unicinctus sperm and spermatogenic cells prepared from the testis are investigated to identify $\alpha-tubulin$ of axoneme microtubules using mouse monoclonal $anti-\alpha-tubulin$ as the first Ab and Gold(10nm) conjugated goat anti-mouse IgG as the Ab marker. The Ag-Ab reaction analyzed excellently the localization of $\alpha-tubulin$ and the gold particles incorporated with the proximal and distal centrioles, manchette microtubules, and flagellum. The gold particles can be also observed in the spermatogenic cells while the cells are still in sperm ball which is composed of a somatic cell and spermatogenic cells. The sperm ball is the functional unit of sperm production in U unicinctus testis. The spermatids are developed from the spermatogenic cells in the sperm ball and released into the testis cavity through a cortical cytoplasmic opening. The spermatid architectures are similar with the mature sperm of the testis cavity in aspects of shape of discoid acrosome, degree of nuclear condensation and ring type of mitochondrion. However, the distal centriole connecting with the flagella can be observed from the mature sperm while the both proximal and distal centrioles reveal only in the spermatids. The proximal centriole is directly connected with nuclear outer membrane during the stage of nuclear condensation and oriented perpendicularly to the distal centriole whose axis coinciding with the longitudinal axis of the spermatozoon. There are indications that the distal centriole is intimately associated with the polymerization of the flagellum. The manchette microtubules appear during spermatid development but the mature sperm have round head and no conspicuous middle piece.

• Journal of Life Science
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• v.28 no.8
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• pp.885-891
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• 2018

Clostridium difficile (C. difficile) toxin A is known to cause acute gut inflammation in humans and animals by triggering cytoskeletal disorganization in gut epithelial cells. In human colonocytes, toxin A blocks microtubule assembly by directly increasing the enzymatic activity of histone deacetylase-6 (HDAC-6), a tubulin-specific deacetylase, thereby markedly decreasing tubulin acetylation, which is essential for microtubule assembly. Microtubule assembly dysfunction-associated alterations (i.e., toxin A-exposed gut epithelial cells) are believed to trigger barrier dysfunction and gut inflammation downstream. We recently showed that potassium acetate blocked toxin A-induced microtubule disassembly by inhibiting HDAC-6. Herein, we tested whether acetic acid (AA), another small acetyl residue-containing agent, could block toxin A-induced tubulin deacetylation and subsequent microtubule assembly. Our results revealed that AA treatment increased tubulin acetylation and enhanced microtubule assembly in an HT29 human colonocyte cell line. AA also clearly increased tubulin acetylation in murine colonic explants. Interestingly, the AA treatment also alleviated toxin A-induced tubulin deacetylation and microtubule disassembly, and MTT assays revealed that AA reduced toxin A-induced cell toxicity. Collectively, these results suggest that AA can block the ability of toxin A to cause microtubule disassembly-triggered cytoskeletal disorganization by blocking toxin A-mediated deacetylation of tubulin.

• Natural Product Sciences
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• v.21 no.4
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• pp.282-288
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• 2015

Viriditoxin is a fungal metabolite isolated from Paecilomyces variotii, which was derived from the giant jellyfish Nemopilema nomurai. Viriditoxin was reported to inhibit polymerization of FtsZ, which is a key protein for bacterial cell division and a structural homologue of eukaryotic tubulin. Both tubulin and FtsZ contain a GTP-binding domain, have GTPase activity, assemble into protofilaments, two-dimensional sheets, and protofilament rings, and share substantial structural identities. Accordingly, we hypothesized that viriditoxin may inhibit eukaryotic cell division by inhibiting tubulin polymerization as in the case of bacterial FtsZ inhibition. Docking simulation of viriditoxin to ${\beta}-tubulin$ indicated that it binds to the paclitaxel-binding domain and makes hydrogen bonds with Thr276 and Gly370 in the same manner as paclitaxel. Viriditoxin suppressed growth of A549 human lung cancer cells, and inhibited cell division with G2/M cell cycle arrest, leading to apoptotic cell death.

• Korean Journal of Bronchoesophagology
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• v.5 no.1
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• pp.42-48
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• 1999

Background： This study was undertaken to detect the patterns of ciliogenesis in newborn rats trachea. Materials and Methods： The experimental animals(Sprague-Dawley strain) were divided five groups, one day, two day, three day, five day and seven day newborns as experimental groups. To obtain differential distribution of $\alpha$-tubulins in the ciliated cells and patterns of ciliogenesis, we used immunohistochemical stain method with mouse anti $\alpha$-tubulin monoclonal antibody as the primary antibody and gold particles conjugated goat anti mouse IgG as the secondary antibody. And we observed the specimens by electron microscope (Hitach-600 Model). Results : 1) The distribution of the $\alpha$ -tubulin reactions in apical zone was slightly decreased from three day after birth. 2) From 5th day after birth, the decreasing number of gold particles in intermediate zone was remarkable. 3) On the comparison with the other zones, the number of gold particles in the Golgi zone for seven days showed no statistical difference. Conclusion : The ciliogenesis of the tracheal ciliated cells in early newborn rat were made via centriolar and acentriolar pathways, in late groups, from five day after birth, the major ciliogenesis pattern might be centriolar pathway.

• Applied Microscopy
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• v.26 no.1
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• pp.11-16
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• 1996

This study was designed to observe the ultrastructure of synoviocytes which are concerned with phagocytic function in the knee joint of the human. The synovia were dissected and were fixed for two hours in 0.2% glutaraldehyde and 4% paraformaldehyde solution and processed and finally infused in 2.3 M sucrose and 20% PVP solution. The tissues were cut with the cryoultramicrotome and labelled with primary antibodies (anti-tubulin, anti-vimentin) and secondary antibody-6 nm colloidal gold particles. The tissues were observed under transmission electronmicroscope. The results were followings. 1. In phagocytic synovial cells, the distributions of tubulin were cytoplasm, especially around vacuoles. 2. In phagocytic synovial cells, the distributions of vimentin were cytoplasm. 3. Both tubulin and vimentin were not located inside of vacuoles. On the basis of above findings, it is obvious that the phagocytic functions are concerned with tubulin, and the phagocytic synovial cells contain vimentin.