Tau confers drug stability but not cold stability to microtubules in living cells.
We previously defined two classes of microtubule polymer in the axons of cultured sympathetic neurons that differ in their sensitivity to nocodazole by roughly 35-fold (Baas and Black (1990) J. Cell Biol. 111, 495-509). Here we demonstrate that virtually all of the microtubule polymer in these axons, including the drug-labile polymer, is stable to cold. What factors account for the unique stability properties of axonal microtubules? In the present study, we have focused on the role of tau, a microtubule-associated protein that is highly enriched in the axon, in determining the stability of microtubules to nocodazole and/or cold in living cells. We used a baculovirus vector to express very high levels of tau in insect ovarian Sf9 cells. The cells respond by extending processes that contain dense bundles of microtubules (Knops et al. (1991) J. Cell Biol. 114, 725-734). Cells induced to express tau were treated with either cold or 2 micrograms/ml nocodazole for times ranging from 5 minutes to 6 hours. The results with each treatment were very different from one another. Virtually all of the polymer was depolymerized within the first 30 minutes in cold, while little or no microtubule depolymerization was detected even after 6 hours in nocodazole. Based on these results, we conclude that tau is almost certainly a factor in conferring drug stability to axonal microtubules, but that factors other than or in addition to tau are required to confer cold stability.
Baas
P W
PW
Department of Anatomy, University of Wisconsin Medical School, Madison 53706.
Pienkowski
T P
TP
Cimbalnik
K A
KA
Toyama
K
K
Bakalis
S
S
Ahmad
F J
FJ
Kosik
K S
KS
D000818:Animals; D001369:Axons / Q000502:physiology / Q000648:ultrastructure*; D016367:Baculoviridae; D002460:Cell Line; D002478:Cells, Cultured; D003080:Cold Temperature; D005260:Female; D008854:Microscopy, Electron; D008870:Microtubules / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure*; D009036:Moths; D009474:Neurons / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure*; D015739:Nocodazole / Q000494:pharmacology; D010053:Ovary; D051381:Rats; D014162:Transfection; D016875:tau Proteins / Q000096:biosynthesis / Q000502:physiology*
D016875:tau Proteins; D015739:Nocodazole