J. Biochem, 1996, Vol. 119, No. 4 731-736
© 1996 Japanese Biochemical Society
research-article |
Pore Formation on Proliferating Yeast Saccharomyces cerevisiae Cell Buds by HM-1 Killer Toxin
*Department of Biochemistry, Niigata College of Pharmacy 5-13-2 Kamishinei-cho, Niigata 950-21
Department of Molecular Genetics, Nippon Roche Research Center Kajiwara, Kamakura 247
AGENE Research Institute Kajiwara, Kamakura 247
1 To whom correspondence should be addressed. Tel: +81-25-269-1224, Fax:+81-25-268-1230, E-Mail: komiyam{at}niigata-pharm.ac.jp
The cytocidal effect of HM-1 produced by Hansenula mrahii on yeast Saccharomyces cerevisiae; cells was studied. The HM-1 strongly inhibited the growth ofS. cerevisiae; cells at a low concentration (IC50;; 2.1 x 10–8;M) by reducing the number of viable cells. The killer action of HM-1 was most efficient when cells were actively proliferating. Cells in a resting state were resistant, but they became HM-1-sensitive after about 90 min of culturing at 30°C, concomitantly with the increment of budding index. In association with the reduction of viable cell number, ultraviolet light-absorbing cellular components were discharged from sensitive cells. HM-1 molecules appear to bind to susceptible cells rather loosely since cells incubated with HM-1 were able to proliferate after having been washed. By phase-contrast light microscopy and scanning electron microscopy, discharge of cell material was observed at the budding portions of HM-1-treated cells. Addition of sorbitol to make the culture medium isotonic partially reduced the cell death induced by HM-1. These results suggest that HM-1 acts on the budding region of proliferating yeast cells, resulting in pore formation, leakage of cell material and eventual cell death.
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