Skip Navigation

This Article
Right arrow Print PDF (8114K)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (66)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Beck, J.
Right arrow Articles by Nassal, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Beck, J.
Right arrow Articles by Nassal, M.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 1995, Vol. 23, No. 24 4954-4962
© 1995

Articles

Efficient hammerhead ribozyme-mediated cleavage of the structured heapatitis B virus encapsidation signal in vitro and in cell extracts, but not in intact cells

Jürgen Beck and Michael Nassal*

Zenturm fuür Molekulare Biologie, Universitaät Heidelberg Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany

*To whom correspondence should be addressed

Received October 9, 1995. Accepted November 8, 1995.

Hepatitis B virus (HBV), the causative agent of B-type hepatitis in man, is a small enveloped DNA virus that replicates through reverse transcription of an RNA intermediate, the terminally redundant RNA pregenome. An essential highly conserved cls-element present twice on this RNA is the encapsidation signal {varepsilon}, a stem-loop structure that is critical for pregenome packaging and reverse transcription, {varepsilon} is hence an attractive target for antiviral therapy. Its structure, however, is a potential obstacle to antivirals whose action depends on hybridization, e.g. ribozymes. Here we demonstrate effective In vitro cleavage inside e by hammerhead ribozymes containing flanking sequences complementary to an adjacent less structured region. Upon cc-transfection with a HBV expression construct corresponding ribozymes embedded in a U6 snRNA context led to a significant, though modest, reduction in the steady-state level of HBV pregenomes. Inactive ribozyme mutants revealed that antisense effects contributed substantially to this reduction, however, efficient {varepsilon} cleavage by the intracellulariy expressed ribozymes was observed in Mg2+-supplemented cell lysates. Artificial HBV pregenomes carrying the ribozymes in els and model RNAs lacking all HBV sequences except {varepsilon} exhibited essentially the same behaviour. Hence, neither the absence of co-localization of ribozyme and target nor a viral component, but rather a cellular factors), is responsible for the strikingly different ribozyme activities inside cells and in cellular extracts.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 J Antimicrob ChemotherHome page
P. Karayiannis
Hepatitis B virus: old, new and future approaches to antiviral treatment
J. Antimicrob. Chemother., April 1, 2003; 51(4): 761 - 785.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
L. J. Bergeron and J.-P. Perreault
Development and comparison of procedures for the selection of delta ribozyme cleavage sites within the hepatitis B virus
Nucleic Acids Res., November 1, 2002; 30(21): 4682 - 4691.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. Amarzguioui, G. Brede, E. Babaie, M. Grotli, B. Sproat, and H. Prydz
Secondary structure prediction and in vitro accessibility of mRNA as tools in the selection of target sites for ribozymes
Nucleic Acids Res., November 1, 2000; 28(21): 4113 - 4124.
[Abstract] [Full Text] [PDF]

Home page
 J. Virol.Home page
J. zu Putlitz, Q. Yu, J. M. Burke, and J. R. Wands
Combinatorial Screening and Intracellular Antiviral Activity of Hairpin Ribozymes Directed against Hepatitis B Virus
J. Virol., July 1, 1999; 73(7): 5381 - 5387.
[Abstract] [Full Text]

Home page
 Proc. Natl. Acad. Sci. USAHome page
D. A. Samarsky, G. Ferbeyre, E. Bertrand, R. H. Singer, R. Cedergren, and M. J. Fournier
A small nucleolar RNA:ribozyme hybrid cleaves a nucleolar RNA target in vivo with near-perfect efficiency
PNAS, June 8, 1999; 96(12): 6609 - 6614.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
H. A. James and I. Gibson
The Therapeutic Potential of Ribozymes
Blood, January 15, 1998; 91(2): 371 - 382.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Scherr, M. Grez, A. Ganser, and J. W. Engels
Specific Hammerhead Ribozyme-mediated Cleavage of Mutant N-ras mRNA in Vitro and ex Vivo. OLIGORIBONUCLEOTIDES AS THERAPEUTIC AGENTS
J. Biol. Chem., May 30, 1997; 272(22): 14304 - 14313.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
G. Ferbeyre, J. Bratty, H. Chen, and R. Cedergren
Cell Cycle Arrest Promotes trans-Hammerhead Ribozyme Action in Yeast
J. Biol. Chem., August 9, 1996; 271(32): 19318 - 19323.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
G. Beterams and M. Nassal
Significant Interference with Hepatitis B Virus Replication by a Core-nuclease Fusion Protein
J. Biol. Chem., March 16, 2001; 276(12): 8875 - 8883.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.