Presence of the betaII isotype of tubulin in the nuclei of cultured mesangial cells from rat kidney.
Tubulin has generally been considered to be a cytosolic protein whose only function is to form microtubules. This assumption is supported by a great deal of evidence derived from immunohistochemical studies using antibodies directed against whole tubulin or its component polypeptides alpha- and beta-tubulin. We have re-examined the intracellular distribution of tubulin using monoclonal antibodies specific for the betaI, betaII, betaIII, and betaIV isotypes of beta-tubulin. Our test system is the cultured rat kidney mesangial cell. We have found that betaIII is absent from these cells and that beta1 and betaIV are present in microtubules throughout the cytosol. In contrast, betaII is present largely in the nuclei. Immunoblotting of purified nuclear extracts shows that the betaII-reactive antigen co-migrates with beta-tubulin. Extraction of the cytosol and chromatin suggests that betaII is concentrated in the nucleoli and also in a reticulated network in the rest of the nucleoplasm. An antibody to tyrosinated alpha-tubulin shows that alpha is also present in the nucleoli. Treatment of the cells with fluorescent colchicine shows an accumulation of colchicine in the nucleoli. Finally, fluorescently labeled alphabetaII-tubulin dimers, when microinjected into the cells, enter the nuclei and are concentrated in the nucleoli. These results suggest that the betaII isotype of tubulin is present as an alphabetaII dimer in the nuclei of cultured mesangial cells and suggest the possibility that different tubulin isotypes may have specific functions within the cell.
Walss
C
C
Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284-7760, USA.
Kreisberg
J I
JI
Ludueña
R F
RF
D000818:Animals; D002453:Cell Cycle; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000737:chemistry; D002478:Cells, Cultured; D003078:Colchicine / Q000032:analysis; D005920:Glomerular Mesangium / Q000737:chemistry* / Q000648:ultrastructure; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D008845:Microinjections; D051381:Rats; D017207:Rats, Sprague-Dawley; D014404:Tubulin / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology
D014404:Tubulin; D003078:Colchicine