Distribution and characteristics of betaII tubulin-enriched microtubules in interphase cells.
We have used a polyclonal antibody (Ab196) that specifically recognizes the betaII tubulin isotype to examine the subcellular distribution and properties of microtubules enriched in this isotype. Antibody specificity was tested by a method that involves the analysis of its interaction with individual beta isotypes. Using photoimaging analysis, we observed betaII tubulin-enriched microtubules in the perinuclear region, as well as in the microtubules close to the periphery of interphase cells. The observed sorting of betaII-enriched microtubules together with the reported increased levels of betaII tubulin in taxol-resistant cells (M. Haber et al., 1995, J. Biol. Chem. 270, 31269-31275) prompted us to study the behavior of microtubules enriched in this isotype after different depolymerizing treatments. After cold or nocodazol treatments, betaII-enriched microtubules anchored at the centrosome and at the cell periphery were observed. In addition, cold-resistant microtubules were marked mainly by the specific anti-betaII tubulin antibody but not by anti-acetylated alpha tubulin, suggesting the presence of different stable microtubule subsets enriched in particular tubulin isoforms.
Armas-Portela
R
R
Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM), Centro Nacional de Biotecnologia, CSIC, Madrid, E-28049, Spain. rarmas@cbm.uam.es
Parrales
M A
MA
Albar
J P
JP
Martinez-A
C
C
Avila
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D002451:Cell Compartmentation; D002522:Chlorocebus aethiops; D003080:Cold Temperature; D003593:Cytoplasm / Q000502:physiology; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D007399:Interphase / Q000502:physiology*; D008870:Microtubules / Q000737:chemistry / Q000502:physiology*; D008969:Molecular Sequence Data; D015739:Nocodazole / Q000494:pharmacology; D020033:Protein Isoforms / Q000302:isolation & purification; D014404:Tubulin / Q000276:immunology / Q000302:isolation & purification*; D014709:Vero Cells
D020033:Protein Isoforms; D014404:Tubulin; D015739:Nocodazole