TY - JOUR
T1 - Detection of human βV-tubulin expression in epithelial cancer cell lines by tubulin proteomics
AU - Verdier-Pinard, Pascal
AU - Shahabi, Shohreh
AU - Wang, Fang
AU - Burd, Berta
AU - Xiao, Hui
AU - Goldberg, Gary L.
AU - Orr, George A.
AU - Horwitz, Susan Band
PY - 2005/12/6
Y1 - 2005/12/6
N2 - Tubulin, the constitutive protein of microtubules, is a heterodimeric protein with an α and β subunit, encoded in vertebrates by six and seven different genes, respectively. Each tubulin isotype can be identified by its divergent C-terminal sequence. Nevertheless, two groups of β-tubulin isotypes can be distinguished by sequence alignment; one includes βI-, βII-, βIVa-, and βIVb-tubulin, and the other includes βIII-, βV-, and βVI-tubulin. βIII-tubulin overexpression has been associated with microtubule destabilization and resistance to Taxol. Recent data indicate that mouse βV-tubulin overexpression in CHO cells results in profound microtubule disorganization and dependence of cells on Taxol for growth. Mouse and human βV-tubulin sequences display several differences, such as their respective extreme C-terminus, suggesting that they may have different effects on microtubule stability and different affinities for drugs. When high-resolution isoelectric focusing, in-gel CNBr cleavage, and mass spectrometry were combined, we detected for the first time the βV-tubulin protein in human cell lines and found that it was highly expressed in Hey, an epithelial ovarian cancer cell line. Our data confirm that human and rodent βV-tubulins are distinct and indicate that, regardless of species, βIII- and βV-tubulin may be expressed in a complementary pattern at the protein level. Therefore, both βIII- and βV-tubulin expression levels should be systematically determined to assess the role of differential tubulin isotype expression in the response of tumors to drugs targeting microtubules.
AB - Tubulin, the constitutive protein of microtubules, is a heterodimeric protein with an α and β subunit, encoded in vertebrates by six and seven different genes, respectively. Each tubulin isotype can be identified by its divergent C-terminal sequence. Nevertheless, two groups of β-tubulin isotypes can be distinguished by sequence alignment; one includes βI-, βII-, βIVa-, and βIVb-tubulin, and the other includes βIII-, βV-, and βVI-tubulin. βIII-tubulin overexpression has been associated with microtubule destabilization and resistance to Taxol. Recent data indicate that mouse βV-tubulin overexpression in CHO cells results in profound microtubule disorganization and dependence of cells on Taxol for growth. Mouse and human βV-tubulin sequences display several differences, such as their respective extreme C-terminus, suggesting that they may have different effects on microtubule stability and different affinities for drugs. When high-resolution isoelectric focusing, in-gel CNBr cleavage, and mass spectrometry were combined, we detected for the first time the βV-tubulin protein in human cell lines and found that it was highly expressed in Hey, an epithelial ovarian cancer cell line. Our data confirm that human and rodent βV-tubulins are distinct and indicate that, regardless of species, βIII- and βV-tubulin may be expressed in a complementary pattern at the protein level. Therefore, both βIII- and βV-tubulin expression levels should be systematically determined to assess the role of differential tubulin isotype expression in the response of tumors to drugs targeting microtubules.
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U2 - 10.1021/bi051004p
DO - 10.1021/bi051004p
M3 - Article
C2 - 16313188
AN - SCOPUS:28544436522
SN - 0006-2960
VL - 44
SP - 15858
EP - 15870
JO - Biochemistry
JF - Biochemistry
IS - 48
ER -