RNA editing by T7 RNA polymerase bypasses InDel mutations causing unexpected phenotypic changes.
Nucleic Acids Res
; 43(8): 3950-63, 2015 Apr 30.
Article
em En
| MEDLINE
| ID: mdl-25824942
ABSTRACT
DNA-dependent T7 RNA polymerase (T7 RNAP) is the most powerful tool for both gene expression and in vitro transcription. By using a Next Generation Sequencing (NGS) approach we have analyzed the polymorphism of a T7 RNAP-generated mRNA pool of the mboIIM2 gene. We find that the enzyme displays a relatively high level of template-dependent transcriptional infidelity. The nucleotide misincorporations and multiple insertions in A/T-rich tracts of homopolymers in mRNA (0.20 and 0.089%, respectively) cause epigenetic effects with significant impact on gene expression that is disproportionally high to their frequency of appearance. The sequence-dependent rescue of single and even double InDel frameshifting mutants and wild-type phenotype recovery is observed as a result. As a consequence, a heterogeneous pool of functional and non-functional proteins of almost the same molecular mass is produced where the proteins are indistinguishable from each other upon ordinary analysis. We suggest that transcriptional infidelity as a general feature of the most effective RNAPs may serve to repair and/or modify a protein function, thus increasing the repertoire of phenotypic variants, which in turn has a high evolutionary potential.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteínas Virais
/
RNA Polimerases Dirigidas por DNA
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Edição de RNA
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Mutação INDEL
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article