Microtubules in the fission yeast Schizosaccharomyces pombe contain only the tyrosinated form of alpha-tubulin.
The state of tubulin tyrosination in the fission yeast Schizosaccharomyces pombe was investigated using a combination of indirect immunofluorescence microscopy and Western blotting. Antibodies specific for the tyrosinated form of alpha-tubulin stained all microtubule arrays in wild type cells and recognised the two alpha-tubulin polypeptides in Western blots of cell extracts enriched for tubulin by DEAE-Sephadex chromatography. Antisera that specifically recognised the detyrosinated, glu, form, on the other hand, gave consistently negative results, both in cells undergoing rapid exponential growth and in those allowed to accumulate in stationary phase. Neither the "ageing" of microtubules, by arresting cells at different points (late G1 or G2/M) in the cell division cycle, nor stabilising them, using D2O, lead to any detectable tubulin detryrosination. These results suggest that S. pombe lacks the carboxypeptidase that carries out the tubulin detyrosination reaction. This is the first report of an organism that possesses the correct C-terminal alpha-tubulin sequence yet fails to carry out this post-translational modification. The implication of this novel finding for the biological role of these events is discussed.
Alfa
C E
CE
Department of Biology, University College, London, England.
Hyams
J S
JS
D000595:Amino Acid Sequence; D015153:Blotting, Western; D002453:Cell Cycle / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000737:chemistry*; D005971:Glutamates / Q000032:analysis; D018698:Glutamic Acid; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000737:chemistry*; D008969:Molecular Sequence Data; D012568:Schizosaccharomyces / Q000737:chemistry*; D014404:Tubulin / Q000737:chemistry*; D014443:Tyrosine / Q000032:analysis
D005656:Fungal Proteins; D005971:Glutamates; D014404:Tubulin; D018698:Glutamic Acid; D014443:Tyrosine