Expression of Reticulomyxa filosa alpha- and beta-tubulins in Escherichia coli yields soluble and partially correctly folded material.

Tubulins are highly conserved multidomain proteins that have to interact with eukaryotic chaperonins to gain their correct three-dimensional conformation. The prokaryotic chaperonin system of GroEL/ES is able to generate intermediate folding states but not natively folded tubulin. To create a system for studying these folding intermediates, tubulins from the giant amoeba Reticulomyxa filosa (alpha 2- and beta 2-tubulin) were expressed in Escherichia coli singly or in tandem.

Dictyostelium gamma-tubulin: molecular characterization and ultrastructural localization.

The centrosome of Dictyostelium discoideum is a nucleus-associated body consisting of an electron-dense, three-layered core surrounded by an amorphous matrix, the corona. To elucidate the molecular and supramolecular architecture of this unique microtubule-organizing center, we have isolated and sequenced the gene encoding gamma-tubulin and have studied its localization in the Dictyostelium centrosome using immunofluorescence and postembedding immunoelectron microscopy. D.

Expression of Reticulomyxa filosa tubulins in Pichia pastoris: regulation of tubulin pools.

We expressed the alpha2- and beta2-tubulin isoforms of the giant freshwater amoeba Reticulomyxa filosa in the methylotrophic yeast Pichia pastoris. Single expression lead to little or no detectable material. Coexpression of both tubulins, however, resulted in a significant increase of expressed proteins.

Primary structure of Neurospora crassa gamma-tubulin.

We report the isolation and characterization of a cDNA-clone for gamma-tubulin from N. crassa. The deduced amino-acid sequence of the cDNA-clone most resembles that of gamma-tubulins from other fungi.