Javascript must be enabled to continue!
Resolution of ribosomal stalling by ABCF ATPases YfmR and YkpA/YbiT
View through CrossRef
ABSTRACT
Efficiency of protein synthesis on the ribosome is strongly affected by the amino acid composition of the assembled amino acid chain. Challenging sequences include proline-rich motifs as well as highly positively and negatively charged amino acid stretches. Members of the F subfamily of ABC ATPases (ABCFs) have been long hypothesised to promote translation of such problematic motifs. In this study we have applied genetics and reporter-based assays to characterise the four housekeeping ABCF ATPases of
Bacillus subtilis
: YdiF, YfmM, YfmR/Uup and YkpA/YbiT. We show that YfmR cooperates with the translation factor EF-P that promotes translation of Pro-rich motifs. Simultaneous loss of both YfmR and EF-P results in a dramatic growth defect. Surprisingly, this growth defect can be largely suppressed though overexpression of an EF-P variant lacking the otherwise crucial 5-amino-pentanolylated residue K32. Using
in vivo
reporter assays, we show that overexpression of YfmR can alleviate ribosomal stalling on Asp-Pro motifs. Finally, we demonstrate that YkpA/YbiT promotes translation of positively and negatively charged motifs but is inactive in resolving ribosomal stalls on proline-rich stretches. Collectively, our results provide insights into the function of ABCF translation factors in modulating protein synthesis in
B. subtilis
.
Title: Resolution of ribosomal stalling by ABCF ATPases YfmR and YkpA/YbiT
Description:
ABSTRACT
Efficiency of protein synthesis on the ribosome is strongly affected by the amino acid composition of the assembled amino acid chain.
Challenging sequences include proline-rich motifs as well as highly positively and negatively charged amino acid stretches.
Members of the F subfamily of ABC ATPases (ABCFs) have been long hypothesised to promote translation of such problematic motifs.
In this study we have applied genetics and reporter-based assays to characterise the four housekeeping ABCF ATPases of
Bacillus subtilis
: YdiF, YfmM, YfmR/Uup and YkpA/YbiT.
We show that YfmR cooperates with the translation factor EF-P that promotes translation of Pro-rich motifs.
Simultaneous loss of both YfmR and EF-P results in a dramatic growth defect.
Surprisingly, this growth defect can be largely suppressed though overexpression of an EF-P variant lacking the otherwise crucial 5-amino-pentanolylated residue K32.
Using
in vivo
reporter assays, we show that overexpression of YfmR can alleviate ribosomal stalling on Asp-Pro motifs.
Finally, we demonstrate that YkpA/YbiT promotes translation of positively and negatively charged motifs but is inactive in resolving ribosomal stalls on proline-rich stretches.
Collectively, our results provide insights into the function of ABCF translation factors in modulating protein synthesis in
B.
subtilis
.
Related Results
ABCF ATPases involved in protein synthesis, ribosome assembly and antibiotic resistance: structural and functional diversification across the tree of life
ABCF ATPases involved in protein synthesis, ribosome assembly and antibiotic resistance: structural and functional diversification across the tree of life
AbstractWithin the larger ABC superfamily of ATPases, ABCF family members eEF3 inSaccharomyces cerevisiaeand EttA inEscherichia colihave been found to function as ribosomal transla...
Beyond self-resistance: ABCF ATPase LmrC is a signal-transducing component of an antibiotic-driven signaling cascade hastening the onset of lincomycin biosynthesis
Beyond self-resistance: ABCF ATPase LmrC is a signal-transducing component of an antibiotic-driven signaling cascade hastening the onset of lincomycin biosynthesis
Abstract
In natural environments, antibiotics are an important instrument of inter-species competition. At subinhibitory concentrations, they act...
Ribosome stalling facilitates chloroplast targeting of nuclear-encoded proteins
Ribosome stalling facilitates chloroplast targeting of nuclear-encoded proteins
Abstract
Ribosomes translate mRNAs with variable elongation rates and frequently undergo transient stalling. Although ribosome stalling is known ...
Introducing ribosomal tandem repeat barcoding for fungi
Introducing ribosomal tandem repeat barcoding for fungi
Abstract
Sequence analysis of the various ribosomal genetic markers is the dominant molecular method for identification and description of fungi. However, there is ...
Ribosomal proteins in halobacteria
Ribosomal proteins in halobacteria
The amino acid sequences of 16 ribosomal proteins from archaebacterium Halobacterium marismortui have been determined by a direct protein chemical method. In addition, amino acid s...
Proton translocation driven by ATP hydrolysis in V‐ATPases
Proton translocation driven by ATP hydrolysis in V‐ATPases
The vacuolar H+‐ATPases (or V‐ATPases) are a family of ATP‐dependent proton pumps responsible for acidification of intracellular compartments and, in certain cases, proton transpor...
Biochemical characterization of P-type copper ATPases
Biochemical characterization of P-type copper ATPases
Copper ATPases, in analogy with other members of the P-ATPase superfamily, contain a catalytic headpiece including an aspartate residue reacting with ATP to form a phosphoenzyme in...
Delivering bad news in emergency care medicine
Delivering bad news in emergency care medicine
Forecasting is a strategy for delivering bad news and is compared to two other strategies, stalling and being blunt. Forecasting provides some warning that bad news is forthcoming ...

