Khoshnan A, Alderete J F
Department of Microbiology, University of Texas Health Science Center, San Antonio 78284-7758.
J Virol. 1993 Dec;67(12):6950-5. doi: 10.1128/JVI.67.12.6950-6955.1993.
Previous studies demonstrated that some isolates of the sexually transmitted protozoan Trichomonas vaginalis are infected with a nonsegmented, double-stranded RNA (dsRNA) virus. A reexamination of the total dsRNA extracted from several virus-harboring isolates indicated the presence of at least three dsRNAs with sizes ranging from 4.8 to 4.3 kbp. The double-stranded nature of each of the three segments was determined by hybridization experiments using riboprobes of opposite polarities obtained from cDNA generated to each of the segments. All three segments were present in agar clones originating from single organisms of T. vaginalis isolates, suggesting that the three segments were not the result of a mixed population of trichomonads harboring different sizes of dsRNA. The three segments were associated with CsCl-purified virus particles, as evidenced by electron microscopy, and RNAse treatment of the preparation containing virus particles did not destroy the dsRNAs. Finally, the individual dsRNA segments were purified for use as probes to determine whether the three dsRNAs shared any sequence homology. Each end-labeled dsRNA segment did not cross-hybridize to any of the other two segments, a finding consistent with the hybridization of labeled cDNAs to only the segments from which they were derived. These results show that the coding capacity of the dsRNA virus may be at least three times greater than that estimated earlier and illustrates further the complexity of this virus-parasite interrelationship.
先前的研究表明,性传播原生动物阴道毛滴虫的一些分离株感染了一种无节段的双链RNA(dsRNA)病毒。对从几种携带病毒的分离株中提取的总dsRNA进行重新检测,结果表明存在至少三种大小在4.8至4.3kbp之间的dsRNA。通过使用从每个片段生成的cDNA获得的极性相反的核糖探针进行杂交实验,确定了这三个片段中每一个的双链性质。所有三个片段都存在于源自阴道毛滴虫分离株单个生物体的琼脂克隆中,这表明这三个片段不是含有不同大小dsRNA的毛滴虫混合群体的结果。如电子显微镜所示,这三个片段与通过CsCl纯化的病毒颗粒相关,并且对含有病毒颗粒的制剂进行RNA酶处理不会破坏dsRNA。最后,对各个dsRNA片段进行纯化,用作探针以确定这三种dsRNA是否具有任何序列同源性。每个末端标记的dsRNA片段都不会与其他两个片段中的任何一个发生交叉杂交,这一发现与标记的cDNA仅与它们所衍生的片段杂交的情况一致。这些结果表明,dsRNA病毒的编码能力可能比先前估计的至少大三倍,并进一步说明了这种病毒与寄生虫相互关系的复杂性。