Identification of a highly conserved sequence element at the 3' terminus of hepatitis C virus genome RNA

被引:344
作者
Kolykhalov, AA
Feinstone, SM
Rice, CM
机构
[1] WASHINGTON UNIV,SCH MED,DEPT MOLEC MICROBIOL,ST LOUIS,MO 63110
[2] US FDA,CTR BIOL EVALUAT & RES,DIV VIROL,BETHESDA,MD 20892
关键词
D O I
10.1128/JVI.70.6.3363-3371.1996
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Previous reports suggest that the hepatitis C virus (HCV) genome RNA terminates with homopolymer tracts of either poly(U) or poly(A), By ligation of synthetic oligonucleotides followed by reverse transcription-PCR, cDNA cloning, and sequence analysis, we determined the 3'-terminal sequence of HCV genome RNA, Our results show that the HCV 3' nontranslated region consists of four elements (positive sense, 5' to 3'): (i) a short sequence with significant variability among genotypes, (ii) a homopolymeric poly(U) tract, (iii) a polypyrimidine stretch consisting of mainly U with interspersed C residues, (iv) a novel sequence of 98 bases, This latter nucleotide sequence is not present in human genomic DNA and is highly conserved among HCV genotypes, The 3'-terminal 46 bases are predicted to form a stable stem-loop structure, Using a quantitative competitive reverse transcription-PCR assay, we show that a substantial fraction of HCV genome RNAs from a high-specific-infectivity inoculum contain this 3'-terminal sequence element, These results indicate that the HCV genome RNA terminates with a highly conserved RNA element which is likely to be required for authentic HCV replication and recovery of infectious RNA from cDNA.
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页码:3363 / 3371
页数:9
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