PXD044929

PXD044929 is an original dataset announced via ProteomeXchange.

Title	Decrypting the functional design of unmodified translation elongation factor P
Description	Stalling of ribosomes due to consecutive proline motifs during polypeptide synthesis is a challenge faced by organisms across all kingdoms. To overcome this, bacteria employ translation elongation factor P (EF-P), while archaea and eukaryotes rely on a/eIF5A. Typically, these elongation factors become active only after undergoing post-translational modifications (PTMs) such as ß-lysinylation, (deoxy-)hypusinylation, rhamnosylation, or 5-aminopentanolyation. An exception to this rule is found in EF-P members of the PGKGP-subfamily, which remain unmodified. However, the mechanism behind the ability of certain bacteria to bypass metabolically and energetically costly PTMs, thus retaining active EF-P, remains unclear. In this study, we investigated the design principles governing the full functionality of unmodified EF-Ps in Escherichia coli. We first screened for naturally unmodified EF-Ps that are active in an E. coli reporter strain. We identified EF-P from Rhodomicrobium vannielii capable of rescuing the growth deficiencies and changes in the proteome of E. coli ΔepmA mutant lacking the gene for the modifying EF-P-(R)-ß-lysine ligase. We then identified specific amino acids in domain I of the unmodified EF-P variant that affected its activity. Ultimately, we transferred these functional properties to other marginally active members of the PGKGP EF-P subfamily, resulting in fully functional unmodified variants in E. coli. These results have not only implications for the improved heterologous expression of polyproline-containing proteins in E. coli but offer applications for other bacterial hosts. Understanding the mechanisms that underlie the functionality of unmodified EF-P provides insights into cellular adaptations to optimize protein synthesis.
HostingRepository	PRIDE
AnnounceDate	2024-04-08
AnnouncementXML	Submission_2024-04-08_13:22:58.384.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Pavel Kielkowski
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Weeksella virosa; NCBI TaxID: 1014; scientific name: Rhodomicrobium vannielii ATCC 17100; NCBI TaxID: 648757;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive HF; Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2023-08-29 04:03:12	ID requested
⏵ 1	2024-04-08 13:22:58	announced

10.1016/J.CELREP.2024.114063;

submitter keyword: post-translational modification,translation elongation factor, ribosome stalling, polyproline-containing proteins

Prof. Dr. Kirsten Jung
contact affiliation	Ludwig-Maximilians-Universität München Biozentrum, Mikrobiologie Großhaderner Str. 2-4 82152 Martinsried
contact email	jung@lmu.de
lab head
Pavel Kielkowski
contact affiliation	LMU Munich
contact email	pavel.kielkowski@cup.lmu.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD044929
     1. Label: PRIDE project
     2. Name: Decrypting the functional design of unmodified translation elongation factor P