Detyrosination of tubulin is not correlated to cold-adaptation of microtubules in cultured cells from the Atlantic cod (Gadus morhua).
Isolated cod brain microtubules from the cold-adapted Atlantic cod (Gadus morhua) have previously been shown to be highly detyrosinated, a post-translational modification of tubulin usually found in stable subsets of microtubules. In this study we found this was not restricted only to isolated brain microtubules. Microtubules in primary cultures of brain and skin cells were composed of both tyrosinated (Tyr)- and detyrosinated (Glu)-tubulin seen by immunocytochemistry. Immunoelectron microscopy of isolated microtubules showed that individual microtubules were composed of a mixture of Tyr- and Glu-tubulin. Leukocytes with extending lamellopodia contained only microtubules stained with the antibody against Tyr-tubulin, and isolated heart tubulin lacked both Tyr- and Glu-tubulin, suggesting that a relative high level of detyrosination is a characteristic of most, but not all, cod microtubules. Brain cell microtubules were more resistant to mitotic inhibitors than skin cell microtubules, but this was not correlated to a difference in detyrosination. Brain and skin cell microtubules were only partially disassembled when incubated at 0 degrees C. Upon reassembly of microtubules at 12 degrees C, microtubules were still made of mixtures of Tyr- and Glu-tubulin, indicating that detyrosination of assembled microtubules is rapid and/or that in cod cells in contrast to mammalian cells, Glu-tubulin can reassemble to microtubules. Our data show that most cod microtubules are highly detyrosinated, but this is not the cause of their cold adaptation or drug stability.
Rutberg
M
M
Department of Zoophysiology, Göteborg University, Sweden.
Modig
C
C
Wallin
M
M
D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000648:ultrastructure; D002478:Cells, Cultured; D003080:Cold Temperature; D005399:Fishes; D019084:Fluorescent Antibody Technique, Indirect; D008099:Liver / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D008870:Microtubules / Q000378:metabolism*; D009206:Myocardium / Q000648:ultrastructure; D011499:Protein Processing, Post-Translational; D013116:Spinal Cord / Q000648:ultrastructure; D014404:Tubulin / Q000032:analysis / Q000378:metabolism*
D014404:Tubulin