ISSN:
0886-1544
Keywords:
microtubules
;
transfection
;
hemagglutinin antigen
;
autoregulation
;
Life and Medical Sciences
;
Cell & Developmental Biology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Medicine
Notes:
A Chinese hamster β-tubulin cDNA, engineered to express a 9 amino acid epitope from the influenza hemagglutinin antigen (HA), was transfected into Chinese hamster ovary (CHO) cells. The recombinant protein (HAβ1-tubulin) appeared to behave normally by the following criteria: immunofluorescence indicated that HAβ1-tubulin incorporated into all classes of interphase and spindle microtubules as well as microtubule organizing centers. The sensitivity of the cells expressing HAβ1-tubulin to Colcemid and taxol was unchanged. A 210 kD microtubule associated protein (MAP) remained associated with microtubules that incorporate HAβ1-tubulin. The synthesis of both endogenous β-tubulin and HAβ1-tubulin was repressed by colchicine. The HAβ1-tubulin incorporated into microtubules to the same extent as the endogenous β-tubulin, and the overall extent of microtubule assembly in transfected cells was unchanged. Finally, trasfected cells had normal growth rates and morphologies. When effects on endogenous tubulin production were measured, it was found that expression of the HAβ1-tubulin reduced the synthesis of endogenous wild-type β-tubulin but increased the synthesis of α-tubulin. At steady state, a small increase in total tubulin consistent with the increased synthesis of α-tubulin was found. The results indicate that expression of excess exogenous β-tubulin perturbs the synthesis of endogenous α-tubulin in a manner that is not easily explained by current models of tubulin regulation. The changes in tubulin synthesis along with degradation of excess tubulin subunits may reflect mechanisms that exist to ensure coordinate levels of α- and β-tubulin for assembly. © 1995 Wiley-Liss, Inc.
Additional Material:
7 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/cm.970310403
