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Molecular and structural characterization of the spindle pole bodies in the fission yeast Schizosaccharomyces japonicus var. japonicus
Author(s) -
Horio Tetsuya,
Kimura Nachi,
Basaki Akemi,
Tanaka Yasuko,
Noguchi Tetsuko,
Akashi Tomohiro,
Tanaka Kenji
Publication year - 2002
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.921
Subject(s) - biology , spindle pole body , schizosaccharomyces pombe , centrosome , mitosis , microbiology and biotechnology , schizosaccharomyces , microtubule , spindle apparatus , microtubule organizing center , microtubule nucleation , cell cycle , cell division , genetics , gene , cell , saccharomyces cerevisiae
The structure and localization of the microtubule organization centres (MTOCs) of the fission yeast Schizosaccharomyces japonicus var. japonicus were examined by fluorescence microscopy and electron microscopy. Spindle pole bodies (SPBs), which are the fungal equivalent of centrosomes, of Sz. japonicus were visualized by immunofluorescent staining using a monoclonal anti‐γ‐tubulin antibody. The behaviour of the SPBs during the cell cycle mostly coincided with previous reports on the most widely used fission yeast Schizosaccharomyces pombe . We cloned the γ‐tubulin gene from Sz. japonicus by PCR using redundant sets of primers corresponding to conserved regions of known γ‐tubulins. The predicted amino acid sequence of Sz. japonicus γ‐tubulin was most similar to the Sz. pombe γ‐tubulin. Under the electron microscope, the SPBs of Sz. japonicus were detected as electron‐dense multilayered structures located just outside the nuclear envelope. The SPBs of Sz. japonicus were composed of three electron‐dense layers and were surrounded by fuzzy material. Each layer showed structural changes according to the progression of the cell cycle. In mitotic cells, the SPBs were located on the fenestrae of the nuclear envelopes through which the mitotic spindle microtubules ran into the nucleoplasm. Our results show that Sz. japonicus is a very potent and attractive organism for the investigation of the microtubule nucleation system and morphogenesis in yeasts. The Accession No. for the nucleotide sequence of the Sz. japonicus gtb1 + gene is AF159163. Copyright © 2002 John Wiley & Sons, Ltd.