Nesterova G F
Genetika. 1993 Apr;29(4):581-603.
Six types of the antagonistic activity (AA) in Saccharomyces were distinguished and characterized. The K1, K2, K3 and K6 activities were associated with the presence of the two kinds of cytoplasmic killer virus-like particles (VLP)--cytoplasmic double-stranded RNAs (dsRNAs) consisting of the main (L) and minor (M) species. All four antagonistic groups contained the L dsRNAs of the same size. The size of the M dsRNAs are dependent on the origin of the host strain. The cytoplasmic inheritance of the K4 activity was observed among the strains which contained no dsRNAs. The AA of this type was not connected with the integrity of mitochondria, also. The K5 activity was under chromosomal control. The optimum and the limits of pH for detection of the AA were dependent on the type of activity. The natural route of extracellular infection by the killer VLPs was the penetration of the VLP into the early protoplast-like spore sprouts. The variability of the K2 activity was studied through mutagenesis of the marked genetic stocks. Twenty six chromosomal genes were involved in the control of this activity. Most of these genetic determinants differed from the chromosomal determinants of the K1 activity in their phenotypes or localization. Four phenotypically different mutational defects of the M-2 plasmid affected the manifestation of the K2 character. Epistatic and cumulative interactions between mutant chromosomal genes controlling the reproduction of the K2 plasmids were observed. Having compared the peculiarities of genetic control of the mutant phenotypes of the K1 and the K2 killers, we acquired the ability to propose a functional model of regulation of the killer virus-like plasmids reproduction. This model defines the interactions between hypothetical gene products and killer dsRNAs or their intermediates. The perspectives of selection of dsRNAs producers were studied. The methods for construction and maintaining the productive strains from the diploids homozygous for a chromosomal mutation increasing the copy number of both K2 killer plasmids were developed. These methods assisted in obtaining the producers suitable for large-scale cultivation.
区分并鉴定了酿酒酵母中的六种拮抗活性(AA)类型。K1、K2、K3和K6活性与两种细胞质杀伤病毒样颗粒(VLP)的存在相关——由主要(L)和次要(M)种类组成的细胞质双链RNA(dsRNA)。所有四个拮抗组都含有相同大小的L dsRNA。M dsRNA的大小取决于宿主菌株的来源。在不含dsRNA的菌株中观察到K4活性的细胞质遗传。这种类型的AA也与线粒体的完整性无关。K5活性受染色体控制。检测AA的最佳pH值和pH范围取决于活性类型。杀伤VLP的细胞外感染自然途径是VLP穿透早期原生质体样孢子芽。通过对标记遗传株系进行诱变研究了K2活性的变异性。26个染色体基因参与了该活性的控制。这些遗传决定因素中的大多数在表型或定位上与K1活性的染色体决定因素不同。M - 2质粒的四个表型不同的突变缺陷影响了K2特性的表现。观察到控制K2质粒复制的突变染色体基因之间的上位性和累积相互作用。比较了K1和K2杀伤者突变表型的遗传控制特性后,我们能够提出杀伤病毒样质粒复制调控的功能模型。该模型定义了假设基因产物与杀伤dsRNA或其中间体之间的相互作用。研究了dsRNA生产者的选择前景。开发了从纯合染色体突变体二倍体构建和维持高产菌株的方法,该突变增加了两种K2杀伤质粒的拷贝数。这些方法有助于获得适合大规模培养的生产者。