An interactive gene network for securin-separase, condensin, cohesin, Dis1/Mtc1 and histones constructed by mass transformation.
The small genome of fission yeast Schizosaccharomyces pombe contains 4824 predicted genes and gene disruption suggests that approximately 850 are essential for viability. To obtain information on interactions among genes required for chromosome segregation, an approach called Strategy B was taken using mass transformation of the 1015 temperature-sensitive (ts) mutants that were made by random mutagenesis and transformed by plasmids carrying the genes for securin, separase, condensin, cohesin, kinetochore microtubule-binding proteins Dis1/Mtc1 or histones. Mutant strains whose phenotypes were either suppressed or inhibited by plasmids were selected. Each plasmid interacted positively or negatively with the average 14 strains. Identification of the mutant gene products by cloning revealed many hitherto unknown interactions. The interactive networks of segregation therefore may consist of genes with a variety of functions. For example, separase/Cut1 interacts with Cdc48/p97/VCP, which stabilizes securin and separase. Surprisingly, S. pombe cdc48 mutants displayed the mitotic phenotype highly similar to separase/cut1 mutants. This approach also provides a novel way of mutant isolation, resulting in two histone H2B strains and a cohesion mutant with a new phenotype.
Yuasa
Tatsuro
T
Department of Biophysics, Graduate School of Science, Kyoto University, Yoshida-Honmachi, Sakyo-ku, Kyoto 606-8501, Japan.
Hayashi
Takeshi
T
Ikai
Nobuyasu
N
Katayama
Toshiaki
T
Aoki
Keita
K
Obara
Takayuki
T
Toyoda
Yusuke
Y
Maruyama
Takeshi
T
Kitagawa
Daisuke
D
Takahashi
Kohta
K
Nagao
Koji
K
Nakaseko
Yukinobu
Y
Yanagida
Mitsuhiro
M
D000251:Adenosine Triphosphatases / Q000235:genetics*; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000235:genetics*; D002868:Chromosomal Proteins, Non-Histone; D015825:Chromosomes, Fungal; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000235:genetics*; D010450:Endopeptidases / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D006657:Histones / Q000235:genetics*; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D046912:Multiprotein Complexes; D009687:Nuclear Proteins / Q000235:genetics*; D010957:Plasmids; D012568:Schizosaccharomyces / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics*; D064247:Separase; D000097722:Cohesins
D000251:Adenosine Triphosphatases; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D017931:DNA Primers; D004268:DNA-Binding Proteins; D010450:Endopeptidases; D005656:Fungal Proteins; D006657:Histones; D008869:Microtubule-Associated Proteins; D046912:Multiprotein Complexes; D009687:Nuclear Proteins; D029702:Schizosaccharomyces pombe Proteins; D064247:Separase; D000097722:Cohesins; C108312:condensin complexes; C094879:dis1 protein, S pombe