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Journal of Virology, June 2000, p. 5502-5508, Vol. 74, No. 12
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Evidence that the 5'-End Cap Structure Is Essential for Encapsidation of Hepatitis B Virus Pregenomic RNA

Jong-Keun Jeong, Gye-Soon Yoon, and Wang-Shick Ryu*

Department of Biochemistry and Institute of Bioscience and Biotechnology, Yonsei University, Seoul, Korea 120-749

Received 9 February 2000/Accepted 21 March 2000

Hepatitis B virus (HBV) replicates by reverse transcription of an RNA intermediate, the pregenomic RNA. The first step of HBV genome replication is the encapsidation of the pregenomic RNA encoding the encapsidation signal, termed varepsilon , into the core particles, which is preceded by recognition and binding of HBV DNA polymerase to varepsilon . The pregenomic RNA contains two identical varepsilon  elements due to its terminal redundancy: one near the 5' end and another near the 3' end. Despite the fact that both varepsilon  elements have an identical sequence, only the 5' varepsilon , but not the 3' varepsilon , is functional for encapsidation. To understand the molecular nature of this position effect, we made a series of lacZ RNA expression plasmids which contain the varepsilon  element at various positions from the 5' end of the transcripts. Following transfection, the lacZ RNAs in cytoplasmic core particles were measured by RNase protection assay for encapsidation. The results indicated that the lacZ RNAs with varepsilon  positioned up to 65 nucleotides from the 5' end were encapsidated, whereas the lacZ RNAs with varepsilon  positioned further downstream were not. Interestingly, the cap-free lacZ RNA transcribed by T7 RNA polymerase was not encapsidated, implying that the 5' cap structure is required for encapsidation of the pregenomic RNA. We hypothesized that HBV DNA polymerase must somehow recognize the cap structure and/or its associated factors, as well as the 5' varepsilon , for encapsidation to occur.

* Corresponding author. Mailing address: 134 Shinchondong, Seodaemungu, Seoul, Korea 120-749. Phone: 82-2-361-2708. Fax: 82-2-362-9897. E-mail: wsryu{at}yonsei.ac.kr.

Journal of Virology, June 2000, p. 5502-5508, Vol. 74, No. 12
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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