Agranovsky A A, Karasev A V, Novikov V K, Lunina N A, Loginov S, Tyulkina L G
A. N. Belozersky Laboratory, Moscow State University, Russia.
J Gen Virol. 1992 Aug;73 ( Pt 8):2085-92. doi: 10.1099/0022-1317-73-8-2085.
RNA from the Hungarian isolate of poa semilatent virus (PSLV) directed in vitro synthesis of 120K, 75K, 25K (coat protein) and 20K polypeptides. In vitro translation of PSLV RNA was blocked by the cap analogue, m7Gpp, thus suggesting that the virus RNA was capped. PSLV RNA could be aminoacylated with [14C]tyrosine in vitro. The sequence of 1.5 kb from the 3' end of the PSLV RNA gamma component revealed two open reading frames (ORFs) separated by a uridine-rich intergenic region. The putative product of the incomplete 5'-proximal ORF showed a close amino acid sequence similarity with the C-terminal segment of the gamma a protein (putative RNA replicase) encoded in the barley stripe mosaic virus (BSMV) RNA gamma, and the 20K product of the 3'-proximal ORF was found to be related to the 17K gamma b product of BSMV. The sequence of 0.8 kb from the 3' end of PSLV RNA beta encompassed two (incomplete) overlapping ORFs whose putative products are related to the beta c and beta d proteins encoded in the similarly arranged ORFs of BSMV RNA beta. Nucleotide sequence homology between the respective parts of the two hordeivirus genomes was restricted to the ORF for gamma a, the spacer between the ORFs for gamma a and gamma b, and the 3' non-coding region, particularly the 95 nucleotide segment at the 3' end representing a tRNA-like structure. Despite limited sequence conservation beyond this segment, the entire 3' non-coding region of PSLV RNA could be folded in a tight pseudoknotted structure closely resembling that of BSMV RNA. Surprisingly, the 'signature' sequence typical for BSMV RNA, internal polydisperse poly(A) intercalated between the coding part of the 3' tRNA-like structure, was not detected in the PSLV genome. Instead, the virus RNA contained several oligoadenylate stretches spaced by other residues, close to the junction of its coding and 3' non-coding portions.
来自匈牙利大麦半潜病毒(PSLV)分离株的RNA在体外指导合成了120K、75K、25K(衣壳蛋白)和20K多肽。PSLV RNA的体外翻译被帽类似物m7Gpp阻断,因此表明病毒RNA被加帽。PSLV RNA在体外可用[14C]酪氨酸进行氨酰化。PSLV RNAγ组分3'端1.5 kb的序列揭示了两个开放阅读框(ORF),由富含尿苷的基因间隔区隔开。不完全的5'-近端ORF的推定产物与大麦条纹花叶病毒(BSMV)RNAγ中编码的γa蛋白(推定的RNA复制酶)的C末端片段显示出紧密的氨基酸序列相似性,并且发现3'-近端ORF的20K产物与BSMV的17Kγb产物相关。PSLV RNAβ3'端0.8 kb的序列包含两个(不完全)重叠的ORF,其推定产物与BSMV RNAβ中类似排列的ORF中编码的βc和βd蛋白相关。两种大麦病毒基因组各部分之间的核苷酸序列同源性仅限于γa的ORF、γa和γb的ORF之间的间隔区以及3'非编码区,特别是3'端代表tRNA样结构的95个核苷酸片段。尽管该片段之外的序列保守性有限,但PSLV RNA的整个3'非编码区可以折叠成紧密的假结结构,与BSMV RNA的结构非常相似。令人惊讶的是,在PSLV基因组中未检测到BSMV RNA典型的“特征”序列,即在3' tRNA样结构的编码部分之间插入的内部多分散多聚腺苷酸。相反,病毒RNA在其编码和3'非编码部分的交界处附近含有几个由其他残基隔开的寡腺苷酸片段。