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Resiliance and proteome response of Escherichia coli to high levels of isoleucine mistranslation

ABSTRACT: Correct pairing of amino acids and tRNA is prerequisite for correct translation of genetic information during protein biosynthesis. Here, we present effects of proteome-wide isoleucine mistranslation in Escherichia coli induced by editing-defective isoleucyl-tRNA synthetase (IleRS). Two types of mistranslation were investigated: substitution of isoleucine (Ile) with either canonical (valine) or noncanonical (norvaline) amino acid. The effects of Ile to Val and Ile to Nva mistranslation were explored on proteome level. High level of Ile substitutions were achieved in editing-deficient strain, leading to excessive protein aggregation. Protein aggregates were analized by LC-MS/MS. Also, interaction of mistranslated proteins with DnaK was explored. The DnaK clients were isolated by pull-down, using endogenously expressed His-tagged DnaK.

ORGANISM(S): Escherichia Coli

SUBMITTER: Ita Gruic Sovulj

PROVIDER: PXD041341 | JPOST Repository | Mon Feb 26 00:00:00 GMT 2024

REPOSITORIES: jPOST

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Resilience and proteome response of Escherichia coli to high levels of isoleucine mistranslation.

Pranjic Marija M Spät Philipp P Semanjski Curkovic Maja M Macek Boris B Gruic-Sovulj Ita I Mocibob Marko M

International journal of biological macromolecules 20240208 Pt 1

Accurate pairing of amino acids and tRNAs is a prerequisite for faithful translation of genetic information during protein biosynthesis. Here we present the effects of proteome-wide mistranslation of isoleucine (Ile) by canonical valine (Val) or non-proteinogenic norvaline (Nva) in a genetically engineered Escherichia coli strain with an editing-defective isoleucyl-tRNA synthetase (IleRS). Editing-defective IleRS efficiently mischarges both Val and Nva to tRNA<sup>Ile</sup> and impairs the trans ...[more]

PMID: 38340920

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