Tags

Type your tag names separated by a space and hit enter

The 3' end of hepatitis E virus (HEV) genome binds specifically to the viral RNA-dependent RNA polymerase (RdRp).
Virology. 2001 Mar 30; 282(1):87-101.V

Abstract

Hepatitis E virus (HEV) is the major cause of acute epidemic and sporadic hepatitis in the developing world. It is a positive-strand RNA virus with a genome length of about 7.2 kb. The replication mechanism of this virus is virtually unexplored. Identification of the regulatory elements involved in initiation of replication may help in designing specific inhibitors for therapy. In the positive-stranded RNA viruses the initiation of replication requires interaction of the 3' end of genome with its RNA-dependent RNA polymerase (RdRp) and possibly host-derived cofactors for synthesis of the minus-strand replicative intermediate. Secondary structure prediction of the conserved 3' end of the infectious HEV genome was carried out to identify possible stem-loop structures necessary for RNA-protein interaction and the model was confirmed by structure probing experiments. Electrophoretic mobility-shift assays showed specific binding of purified and refolded recombinant HEV RdRp protein to the 3' end of its RNA genome containing the poly(A) stretch. Mutations at the 3' end, in which the stem-loop structures were partially or completely destroyed or recreated revealed that the two stem-loop structures SL1 and SL2 at the 3' end and the poly(A) stretch are necessary for this binding. The interacting nucleotides in such an interaction were further identified by generating footprints of the complex by Pb(II)-induced hydrolysis. This specific binding of viral RdRp to the 3' end of HEV RNA directs the synthesis of complementary-strand RNA and thus such a binding domain might assume the role of a possible cis-acting element as a potential site for the initiation of replication.

Authors+Show Affiliations

Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029, India.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

11259193

Citation

Agrawal, S, et al. "The 3' End of Hepatitis E Virus (HEV) Genome Binds Specifically to the Viral RNA-dependent RNA Polymerase (RdRp)." Virology, vol. 282, no. 1, 2001, pp. 87-101.
Agrawal S, Gupta D, Panda SK. The 3' end of hepatitis E virus (HEV) genome binds specifically to the viral RNA-dependent RNA polymerase (RdRp). Virology. 2001;282(1):87-101.
Agrawal, S., Gupta, D., & Panda, S. K. (2001). The 3' end of hepatitis E virus (HEV) genome binds specifically to the viral RNA-dependent RNA polymerase (RdRp). Virology, 282(1), 87-101.
Agrawal S, Gupta D, Panda SK. The 3' End of Hepatitis E Virus (HEV) Genome Binds Specifically to the Viral RNA-dependent RNA Polymerase (RdRp). Virology. 2001 Mar 30;282(1):87-101. PubMed PMID: 11259193.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The 3' end of hepatitis E virus (HEV) genome binds specifically to the viral RNA-dependent RNA polymerase (RdRp). AU - Agrawal,S, AU - Gupta,D, AU - Panda,S K, PY - 2001/3/22/pubmed PY - 2001/5/1/medline PY - 2001/3/22/entrez SP - 87 EP - 101 JF - Virology JO - Virology VL - 282 IS - 1 N2 - Hepatitis E virus (HEV) is the major cause of acute epidemic and sporadic hepatitis in the developing world. It is a positive-strand RNA virus with a genome length of about 7.2 kb. The replication mechanism of this virus is virtually unexplored. Identification of the regulatory elements involved in initiation of replication may help in designing specific inhibitors for therapy. In the positive-stranded RNA viruses the initiation of replication requires interaction of the 3' end of genome with its RNA-dependent RNA polymerase (RdRp) and possibly host-derived cofactors for synthesis of the minus-strand replicative intermediate. Secondary structure prediction of the conserved 3' end of the infectious HEV genome was carried out to identify possible stem-loop structures necessary for RNA-protein interaction and the model was confirmed by structure probing experiments. Electrophoretic mobility-shift assays showed specific binding of purified and refolded recombinant HEV RdRp protein to the 3' end of its RNA genome containing the poly(A) stretch. Mutations at the 3' end, in which the stem-loop structures were partially or completely destroyed or recreated revealed that the two stem-loop structures SL1 and SL2 at the 3' end and the poly(A) stretch are necessary for this binding. The interacting nucleotides in such an interaction were further identified by generating footprints of the complex by Pb(II)-induced hydrolysis. This specific binding of viral RdRp to the 3' end of HEV RNA directs the synthesis of complementary-strand RNA and thus such a binding domain might assume the role of a possible cis-acting element as a potential site for the initiation of replication. SN - 0042-6822 UR - https://www.unboundmedicine.com/medline/citation/11259193/The_3'_end_of_hepatitis_E_virus__HEV__genome_binds_specifically_to_the_viral_RNA_dependent_RNA_polymerase__RdRp__ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0042-6822(00)90819-5 DB - PRIME DP - Unbound Medicine ER -