Nagasaki Keizo, Shirai Yoko, Takao Yoshitake, Mizumoto Hiroyuki, Nishida Kensho, Tomaru Yuji
National Research Institute of Fisheries and Environment of Inland Sea, Fisheries Research Agency, 2-17-5 Maruishi, Hatsukaichi, Hiroshima 739-0452, Japan.
Appl Environ Microbiol. 2005 Dec;71(12):8888-94. doi: 10.1128/AEM.71.12.8888-8894.2005.
Heterocapsa circularisquama RNA virus (HcRNAV) has at least two ecotypes (types UA and CY) that have intraspecies host specificities which are complementary to each other. We determined the complete genomic RNA sequence of two typical HcRNAV strains, HcRNAV34 and HcRNAV109, one of each ecotype. The nucleotide sequences of the viruses were 97.0% similar, and each had two open reading frames (ORFs), ORF-1 coding for a putative polyprotein having protease and RNA-dependent RNA polymerase (RdRp) domains and ORF-2 encoding a single major capsid protein. Phylogenetic analysis of the RdRp amino acid sequence suggested that HcRNAV belongs to a new previously unrecognized virus group. Four regions in ORF-2 had amino acid substitutions when HcRNAV34 was compared to HcRNAV109. We used a reverse transcription-nested PCR system to amplify the corresponding regions and also examined RNAs purified from six other HcRNAV strains with known host ranges. We also looked at natural marine sediment samples. Phylogenetic dendrograms for the amplicons correlated with the intraspecies host specificities of the test virus strains. The cloned sequences found in sediment also exhibited considerable similarities to either the UA-type or CY-type sequence. The tertiary structure of the capsid proteins predicted using computer modeling indicated that many of the amino acid substitutions were located in regions on the outside of the viral capsid proteins. This strongly suggests that the intraspecies host specificity of HcRNAV is determined by nanostructures on the virus surface that may affect binding to suitable host cells. Our study shows that capsid alterations can change the phytoplankton-virus (host-parasite) interactions in marine systems.
圆形异帽藻RNA病毒(HcRNAV)至少有两种生态型(UA型和CY型),它们具有种内宿主特异性,且相互互补。我们测定了两种典型的HcRNAV毒株HcRNAV34和HcRNAV109的完整基因组RNA序列,每种生态型各一个。这两种病毒的核苷酸序列相似度为97.0%,且各有两个开放阅读框(ORF),ORF-1编码一个推定的多蛋白,该多蛋白具有蛋白酶和RNA依赖性RNA聚合酶(RdRp)结构域,ORF-2编码一种主要的衣壳蛋白。对RdRp氨基酸序列的系统发育分析表明,HcRNAV属于一个此前未被识别的新病毒组。与HcRNAV109相比,HcRNAV34的ORF-2中有四个区域存在氨基酸替换。我们使用逆转录巢式PCR系统扩增相应区域,并检测了从其他六种已知宿主范围的HcRNAV毒株中纯化的RNA。我们还研究了天然海洋沉积物样本。扩增子的系统发育树状图与测试病毒株的种内宿主特异性相关。在沉积物中发现的克隆序列与UA型或CY型序列也表现出相当高的相似性。使用计算机建模预测的衣壳蛋白三级结构表明,许多氨基酸替换位于病毒衣壳蛋白外部的区域。这有力地表明,HcRNAV的种内宿主特异性是由病毒表面的纳米结构决定的,这些纳米结构可能会影响与合适宿主细胞的结合。我们的研究表明,衣壳改变可以改变海洋系统中浮游植物与病毒(宿主-寄生虫)的相互作用。