Agris C H, Blake K R, Miller P S, Reddy M P, Ts'o P O
Biochemistry. 1986 Oct 7;25(20):6268-75. doi: 10.1021/bi00368a065.
Oligodeoxyribonucleoside methylphosphonates which have sequences complementary to the initiation codon regions of N, NS, and G vesicular stomatitis virus (VSV) mRNAs were tested for their ability to inhibit translation of VSV mRNA in a cell-free system and in VSV-infected mouse L cells. In a rabbit reticulocyte lysate cell-free system, the oligomers complementary to N (oligomer I) and NS (oligomer II) mRNAs inhibited translation of VSV N and NS mRNAs whereas oligomer III had only a slight inhibitory effect on N protein synthesis. At 100 and 150 microM, oligomer I specifically inhibited N protein synthesis in the lysate. In contrast, at 150 microM, oligomer II inhibited both N and NS protein synthesis. This reduced specificity of inhibition may be due to the formation of partial duplexes between oligomer II and VSV N mRNA. The oligomers had little or no inhibitory effects on the synthesis of globin mRNA in the same lysate system. Oligomers I-III specifically inhibited the synthesis of all five viral proteins in VSV-infected cells in a concentration-dependent manner. The oligomers had no effects on cellular protein synthesis in uninfected cells nor on cell growth. An oligothymidylate which forms only weak duplexes with poly(rA) had just a slight effect on VSV protein synthesis and yield of virus. Oligomers I-III have extensive partial complementarity with the coding regions of L mRNA. The nonspecific inhibition of viral protein synthesis in infected cells may reflect the role of N, NS, and/or L proteins in the replication and transcription of viral RNA or result from duplex formation between the oligomers and complementary, plus-strand viral RNA.(ABSTRACT TRUNCATED AT 250 WORDS)
对具有与水泡性口炎病毒(VSV)N、NS和G mRNA起始密码子区域互补序列的寡脱氧核糖核苷甲基膦酸酯,检测了它们在无细胞系统和VSV感染的小鼠L细胞中抑制VSV mRNA翻译的能力。在兔网织红细胞裂解物无细胞系统中,与N(寡聚物I)和NS(寡聚物II)mRNA互补的寡聚物抑制了VSV N和NS mRNA的翻译,而寡聚物III对N蛋白合成仅有轻微抑制作用。在100和150微摩尔浓度时,寡聚物I特异性抑制裂解物中的N蛋白合成。相反,在150微摩尔浓度时,寡聚物II抑制N和NS蛋白合成。这种抑制特异性的降低可能是由于寡聚物II与VSV N mRNA之间形成了部分双链体。在同一裂解物系统中,这些寡聚物对珠蛋白mRNA的合成几乎没有抑制作用。寡聚物I - III以浓度依赖的方式特异性抑制VSV感染细胞中所有五种病毒蛋白的合成。这些寡聚物对未感染细胞中的细胞蛋白合成以及细胞生长没有影响。仅与聚(rA)形成弱双链体的寡聚胸苷酸对VSV蛋白合成和病毒产量仅有轻微影响。寡聚物I - III与L mRNA的编码区域有广泛的部分互补性。感染细胞中病毒蛋白合成的非特异性抑制可能反映了N、NS和/或L蛋白在病毒RNA复制和转录中的作用,或者是由于寡聚物与互补的正链病毒RNA之间形成双链体所致。(摘要截短于250字)