Detyrosinated (Glu) microtubules are stabilized by an ATP-sensitive plus-end cap.
Many cell types contain a subset of long-lived, 'stable' microtubules that differ from dynamic microtubules in that they are enriched in post-translationally detyrosinated tubulin (Glu-tubulin). Elevated Glu tubulin does not stabilize the microtubules and the mechanism for the stability of Glu microtubules is not known. We used detergent-extracted cell models to investigate the nature of Glu microtubule stability. In these cell models, Glu microtubules did not incorporate exogenously added tubulin subunits on their distal ends, while >70% of the bulk microtubules did. Ca(2+)-generated fragments of Glu microtubules incorporated tubulin, showing that Glu microtubule ends are capped. Consistent with this, Glu microtubules in cell models were resistant to dilution-induced breakdown. Known microtubule end-associated proteins (EB1, APC, p150(Glued) and vinculin focal adhesions) were not localized on Glu microtubule ends. ATP, but not nonhydrolyzable analogues, induced depolymerization of Glu microtubules in cell models. Timelapse and photobleaching studies showed that ATP triggered subunit loss from the plus end. ATP breakdown of Glu microtubules was inhibited by AMP-PNP and vanadate, but not by kinase or other inhibitors. Additional experiments showed that conventional kinesin or kif3 were not involved in Glu microtubule capping. We conclude that Glu microtubules are stabilized by a plus-end cap that includes an ATPase with properties similar to kinesins.
Infante
A S
AS
Integrated Program in Cellular, Molecular and Biophysical Studies and Department of Anatomy and Cell Biology, Columbia University, New York, NY 10032, USA.
Stein
M S
MS
Zhai
Y
Y
Borisy
G G
GG
Gundersen
G G
GG
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D002522:Chlorocebus aethiops; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D004847:Epithelial Cells / Q000166:cytology* / Q000502:physiology* / Q000648:ultrastructure; D018698:Glutamic Acid; D007668:Kidney; D008870:Microtubules / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D011499:Protein Processing, Post-Translational; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism
D014404:Tubulin; D018698:Glutamic Acid; D014443:Tyrosine; D000255:Adenosine Triphosphate; D002118:Calcium