title: RNA editing by T7 RNA polymerase bypasses InDel mutations causing unexpected phenotypic changes.
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.
authors.affiliation: Department of Microbiology, University of Gdansk, Gdansk 80-308, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Wons
authors.affiliation: Department of Microbiology, University of Gdansk, Gdansk 80-308, Poland.
authors.forename: Beata
authors.initials: B
authors.lastname: Furmanek-Blaszk
authors.affiliation: Department of Microbiology, University of Gdansk, Gdansk 80-308, Poland marian.sektas@biol.ug.edu.pl.
authors.forename: Marian
authors.initials: M
authors.lastname: Sektas
mesh_terms: D000596:Amino Acids / Q000032:analysis; D004260:DNA Repair; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D044127:Epigenesis, Genetic; D018965:Frameshifting, Ribosomal; D054643:INDEL Mutation; D008780:Methyltransferases / Q000737:chemistry / Q000235:genetics; D016955:Moraxella bovis / Q000201:enzymology / Q000235:genetics; D010641:Phenotype; D017393:RNA Editing; D012333:RNA, Messenger / Q000737:chemistry; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D012333:RNA, Messenger; D014764:Viral Proteins; D008780:Methyltransferases; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases
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