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麦芽糖假丝酵母中对放线菌酮敏感和耐药核糖体的研究。

Studies on cycloheximide-sensitive and cycloheximide-resistant ribosomes in the yeast Candida maltosa.

作者信息

Mutoh E, Ohta A, Takagi M

机构信息

Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113, Japan.

出版信息

Gene. 1998 Dec 11;224(1-2):9-15. doi: 10.1016/s0378-1119(98)00518-6.

Abstract

Cycloheximide sensitivity or resistance in yeast is under the control of genes encoding different forms of ribosomal protein L41. In our previous studies, we have shown by isolating L41-Q1a, L41-P1a and their respective allelic genes, L41-Q1b and L41-P1b, from the partial diploid genome of C. maltosa, that this species, which is inducibly resistant to CYH, has both types of the L41 genes and that the expression of at least one of the L41-Q genes is induced by CYH, whereas L41-P genes are constitutively expressed. Here, we have identified another L41 (L41-Q2a), its allelic gene (L41-Q2b) and a third gene (L41-Q3) from the genome of C. maltosa. By gene disruption experiments, we now show that L41-Q1a and L41-Q1b are not responsible for the resistance to CYH and that the DeltaL41-Ps strain, which has only functional L41-Q genes, shows constitutive resistance to CYH, but grows more slowly than the DeltaL41-Qs strain, which has only functional L41-P genes, in the absence of CYH. Our results also show that in vitro, ribosomes containing L41-Q-type are less active in translation than those containing L41-P-type, although only the former ribosomes are active in the presence of CYH. These data suggest that ribosomes containing L41-Q-type are less active under normal growth conditions, but that this activity is not affected in the presence of CYH. We discuss the possible multi-step evolutionary event(s) by which C. maltosa has acquired the property of inducible resistance to CYH.

摘要

酵母对放线菌酮的敏感性或抗性受编码不同形式核糖体蛋白L41的基因控制。在我们之前的研究中,通过从麦芽糖假丝酵母的部分二倍体基因组中分离出L41-Q1a、L41-P1a及其各自的等位基因L41-Q1b和L41-P1b,我们发现,这种对放线菌酮具有诱导抗性的物种同时拥有两种类型的L41基因,并且至少一个L41-Q基因的表达受放线菌酮诱导,而L41-P基因则组成性表达。在此,我们从麦芽糖假丝酵母的基因组中鉴定出了另一个L41(L41-Q2a)、其等位基因(L41-Q2b)和第三个基因(L41-Q3)。通过基因敲除实验,我们现在表明L41-Q1a和L41-Q1b与对放线菌酮的抗性无关,并且仅具有功能性L41-Q基因的DeltaL41-Ps菌株对放线菌酮具有组成性抗性,但在没有放线菌酮的情况下,其生长速度比仅具有功能性L41-P基因的DeltaL41-Qs菌株慢。我们的结果还表明,在体外,含有L41-Q型的核糖体在翻译中的活性低于含有L41-P型的核糖体,尽管只有前者的核糖体在放线菌酮存在时具有活性。这些数据表明,含有L41-Q型的核糖体在正常生长条件下活性较低,但这种活性在放线菌酮存在时不受影响。我们讨论了麦芽糖假丝酵母获得对放线菌酮诱导抗性特性可能的多步进化事件。

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