Document Detail

Design of artificial short-chained RNA species that are replicated by Q beta replicase.
MedLine Citation:
PMID:  7530049     Owner:  NLM     Status:  MEDLINE    
Different RNA species that are replicated by Q beta replicase have related secondary structures: for both plus and minus strands, "leader" stem structures were found at their 5' termini, while their 3' termini were unpaired. Parallel structures in complementary strands rather than antiparallel ones require the occurrence of wobble pairs and other imperfections in the stem regions. To test whether the leader structures are required for replication, artificial RNA sequences were synthesized by transcription from synthetic oligodeoxynucleotides with T7 RNA polymerase and assayed for their ability to be replicated by Q beta replicase. A synthetic short RNA species known to be replicated was amplified, forming a stable quasi-species; i.e., its sequence was conserved during hundreds of replication rounds. A synthetic mutant of this sequence that stabilized the leader in one strand but favored a 3'-terminal stem in the other one led to the complete loss of template activity. When new RNA sequences with the described structural requirements were designed and synthesized, their template activity was too low to be directly measurable; however, incubation with replicase produced replicating RNA whose sequence was closely related to the synthesized RNA species. The most likely interpretation is that the designed sequences were in a low montainous region in the replication fitness landscape and were optimized during amplification by Q beta replicase to a nearby fitness peak. The structural features postulated to be required for replication were not only conserved but even improved in the outgrowing mutants.(ABSTRACT TRUNCATED AT 250 WORDS)
H Zamora; R Luce; C K Biebricher
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochemistry     Volume:  34     ISSN:  0006-2960     ISO Abbreviation:  Biochemistry     Publication Date:  1995 Jan 
Date Detail:
Created Date:  1995-02-23     Completed Date:  1995-02-23     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1261-6     Citation Subset:  IM    
Max-Planck-Institute for Biophysical Chemistry, Am Fassberg, Göttingen, Federal Republic of Germany.
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MeSH Terms
Base Sequence
Hydrogen Bonding
Molecular Sequence Data
Nucleic Acid Conformation
Q beta Replicase / metabolism*
RNA / chemistry,  ultrastructure
RNA, Viral / biosynthesis*,  ultrastructure
Structure-Activity Relationship
Templates, Genetic
Reg. No./Substance:
0/RNA, Viral; 63231-63-0/RNA; EC 2.7.7.-/Q beta Replicase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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