Cold-stable microtubules from Antarctic fishes contain unique alpha tubulins.
The cytoplasmic microtubules of Antarctic fishes assemble from their tubulin subunits at physiological body temperatures in the range -2 to +2 degrees C. Our objective is to determine the structural features that enhance the assembly of Antarctic fish tubulins at low temperatures. Here we compare the structures of tubulin subunits from three Antarctic fishes (Notothenia gibberifrons, Notothenia coriiceps neglecta, and Chaenocephalus aceratus), from three temperate fishes (the dogfish shark Mustelus canis, the channel catfish Ictalurus punctatus, and the goosefish Lophius americanus), and from a mammal (the cow Bos taurus). When reduced, carboxymethylated, and examined by polyacrylamide gel electrophoresis, multiple alpha chains were observed in tubulins from the Antarctic fishes, the catfish, and the goosefish; dogfish and bovine alpha tubulins migrated as single components on this gel system. Prominent in the Antarctic fish tubulins was an alpha variant that migrated more rapidly than the bovine alpha chain; smaller amounts of a rapidly migrating alpha chain were also present in catfish and goosefish tubulins. The beta tubulins of the fishes, with the exception of the goosefish, resolved into major and minor variants with mobilities similar to those of beta 1 and beta 2 tubulins from bovine brain. Peptide mapping demonstrated that the alpha tubulins of Antarctic fishes were similar in structure, yet differed from the alpha chains of the dogfish and the cow (which, in turn, were similar to each other). In contrast, the beta tubulins from these organisms gave peptide patterns of near identity. Finally, the alpha chains of native tubulins from N. coriiceps neglecta and the cow differed in the sensitivity of their C-terminal domains to digestion by subtilisin. These results demonstrate that the alpha tubulins of Antarctic fishes (but not their beta chains) differ structurally from those of temperate fishes and a mammal.
D000818:Animals; D000864:Antarctic Regions; D001921:Brain / Q000502:physiology; D003081:Cold Climate; D005399:Fishes / Q000502:physiology*; D008870:Microtubules / Q000032:analysis / Q000648:ultrastructure*; D008970:Molecular Weight; D013045:Species Specificity; D014404:Tubulin / Q000302:isolation & purification / Q000502:physiology*
D014404:Tubulin