Resolution of ribosomal stalling by EF-P and ABCF ATPases YfmR and YkpA/YbiT
(2024) In Nucleic Acids Research p.1-13- 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.... (More)
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.
(Less)
- author
- Takada, Hiraku ; Fujiwara, Keigo ; Atkinson, Gemma C LU ; Chiba, Shinobu and Hauryliuk, Vasili LU
- organization
- publishing date
- 2024-06-29
- type
- Contribution to journal
- publication status
- epub
- subject
- in
- Nucleic Acids Research
- article number
- gkae556
- pages
- 1 - 13
- publisher
- Oxford University Press
- external identifiers
-
- pmid:38943426
- ISSN
- 1362-4962
- DOI
- 10.1093/nar/gkae556
- language
- English
- LU publication?
- yes
- additional info
- © The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.
- id
- d0b367bc-7b2e-423f-a437-93051d6d1462
- date added to LUP
- 2024-07-02 20:47:42
- date last changed
- 2024-07-03 10:17:02
@article{d0b367bc-7b2e-423f-a437-93051d6d1462, abstract = {{<p>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.</p>}}, author = {{Takada, Hiraku and Fujiwara, Keigo and Atkinson, Gemma C and Chiba, Shinobu and Hauryliuk, Vasili}}, issn = {{1362-4962}}, language = {{eng}}, month = {{06}}, pages = {{1--13}}, publisher = {{Oxford University Press}}, series = {{Nucleic Acids Research}}, title = {{Resolution of ribosomal stalling by EF-P and ABCF ATPases YfmR and YkpA/YbiT}}, url = {{http://dx.doi.org/10.1093/nar/gkae556}}, doi = {{10.1093/nar/gkae556}}, year = {{2024}}, }