Preparation of a monoclonal antibody specific for the class I isotype of β-tubulin: The β isotypes of tubulin differ in their cellular distributions within human tissues

Roach, Mary Carmen; Boucher, Virginia; Walss, Consuelo; Ravdin, Peter M.; Ludueña, Richard F.

doi:10.1002/(sici)1097-0169(1998)39:4<273::aid-cm3>3.0.co;2-4

Cited by 66 publications

(18 citation statements)

References 27 publications

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“…Consistent with our results is the finding of both β-tubulin classes I and II by immunostaining in many human tissues and cell types [60]. To understand the relationship between the drug target levels and the development of drug resistance, it is essential to know the actual protein amounts in tissues and cells.…”

Section: Discussionsupporting

confidence: 90%

β class II tubulin predominates in normal and tumor breast tissues

et al. 2003

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Common agents currently used in treating metastatic breast cancer are the antimitotics paclitaxel and docetaxel [1,2]. These drugs bind to β-tubulin, a major protein in mitotic spindles, and halt cell division at metaphase. Their effectiveness in cancer chemotherapy is thought to be due to their ability to reduce the dynamics of microtubules in mitotic spindles, thus preventing spindle assembly and interrupting the normal movement of sister chromatids toward the spindle poles [3][4][5][6][7][8][9]. Tubulin, a 100 kDa αβ heterodimer, is structurally heterogeneous, with seven genes encoding α-tubulin and β-tubulin isotypes. The major differences between isotype classes reside in the last 15-20 amino acids of the carboxy-termini et al., licensee BioMed Central Ltd (Print ISSN 1465-5411; Online ISSN 1465-542X). This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL. AbstractBackground: Antimitotic chemotherapeutic agents target tubulin, the major protein in mitotic spindles. Tubulin isotype composition is thought to be both diagnostic of tumor progression and a determinant of the cellular response to chemotherapy. This implies that there is a difference in isotype composition between normal and tumor tissues.

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Section: Discussionsupporting

confidence: 90%

β class II tubulin predominates in normal and tumor breast tissues

et al. 2003

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“…βI is the most widespread of the vertebrate β isotypes, occurring in almost all tissues (Roach et al 1998); our results presented here suggest that βI is vital for cell viability. Hitherto the only clues to specific functions for βI are that it appears not to interact with actin filaments (Lezama et al 2001) and that, like βIV, it occurs in axonemes, although not as prominently as does βIV (Jensen-Smith et al 2003).…”

Section: Discussionsupporting

confidence: 48%

“…This is not likely to be due to a decrease in total tubulin expression, since βI is not the major isotype expressed in these cells, and a larger degree of suppression of either βII or βIII does not result in any significant loss of viability. The fact that βI is so widespread in cells and tissues (Roach et al 1998) suggests that it is capable of carrying out the canonical microtubule functions of mitosis and intracellular transport. The fact that βI occurs in both cell bodies and neurites is consistent with these roles and may account for its being required for cell viability, especially if βII and βIII have evolved to have more specialized roles.…”

Section: Discussionmentioning

confidence: 99%

“…The monoclonal antibodies specific to the β isotypes of tubulin (βI, βII, βIII) were prepared as previously described (Banerjee et al 1990, 1992, 1988; Roach et al 1998). Hybridoma supernatants containing antibodies to βI (SAP.4G5), βII (JDR.3B8), βIII (SDL.3D10), and βIV (ONS.1A6) were passed through a protein A-agarose column and washed with PBS.…”

Section: Methodsmentioning

confidence: 99%

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The β isotypes of tubulin in neuronal differentiation

2010

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The differences among the vertebrate β isotypes of tubulin are highly conserved in evolution, suggesting that they have functional significance. To address this, we have used differentiating neuroblastoma cells as a model system. These cells express the βI, βII, and βIII isotypes. Although there is no difference prior to differentiation, a striking difference is seen after differentiation. Both βI and βIII occur in cell bodies and neurites, while βII occurs mostly in neurites. Knocking down βI causes a large decrease in cell viability while silencing βII and βIII does not. Knocking down βII causes a large decrease in neurite outgrowth without affecting viability. Knocking down βIII has little effect on neurite outgrowth and only decreases viability if cells are treated with glutamate and glycine, a combination known to generate free radicals and reactive oxygen species. It appears, therefore, that βI is required for cell viability, βII for neurite outgrowth and βIII for protection against free radicals and reactive oxygen species.

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“…In humans, several α and β tubulin isotypes have been identified and characterized (Lu and Luduena, 1994; Luduena, 1998; Roach et al 1998). While α tubulin must play an obvious role in the determination of MT function, we have chosen to focus only on β tubulin for this discussion, as most of the available data in the literature deals with this protein as a target for drug action and protein-protein interactions.…”

Section: Introductionmentioning

confidence: 99%

The Roles of β-Tubulin Mutations and Isotype Expression in Acquired Drug Resistance

et al. 2007

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The antitumor drug paclitaxel stabilizes microtubules and reduces their dynamicity, promoting mitotic arrest and eventually apoptosis. Upon assembly of the α/β-tubulin heterodimer, GTP becomes bound to both the α and β-tubulin monomers. During microtubule assembly, the GTP bound to β-tubulin is hydrolyzed to GDP, eventually reaching steady-state equilibrium between free tubulin dimers and those polymerized into microtubules. Tubulin-binding drugs such as paclitaxel interact with β-tubulin, resulting in the disruption of this equilibrium. In spite of several crystal structures of tubulin, there is little biochemical insight into the mechanism by which anti-tubulin drugs target microtubules and alter their normal behavior. The mechanism of drug action is further complicated, as the description of altered β-tubulin isotype expression and/or mutations in tubulin genes may lead to drug resistance as has been described in the literature. Because of the relationship between β-tubulin isotype expression and mutations within β-tubulin, both leading to resistance, we examined the properties of altered residues within the taxane, colchicine and Vinca binding sites. The amount of data now available, allows us to investigate common patterns that lead to microtubule disruption and may provide a guide to the rational design of novel compounds that can inhibit microtubule dynamics for specific tubulin isotypes or, indeed resistant cell lines. Because of the vast amount of data published to date, we will only provide a broad overview of the mutational results and how these correlate with differences between tubulin isotypes. We also note that clinical studies describe a number of predictive factors for the response to anti-tubulin drugs and attempt to develop an understanding of the features within tubulin that may help explain how they may affect both microtubule assembly and stability.

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Preparation of a monoclonal antibody specific for the class I isotype of β-tubulin: The β isotypes of tubulin differ in their cellular distributions within human tissues

Cited by 66 publications

References 27 publications

β class II tubulin predominates in normal and tumor breast tissues

β class II tubulin predominates in normal and tumor breast tissues

The β isotypes of tubulin in neuronal differentiation

The Roles of β-Tubulin Mutations and Isotype Expression in Acquired Drug Resistance

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