Dataset Information

Mass spectrometry analysis of four translation initiation proteins, EIF4E3, EIF4E4, EIF4G3 and EIF4G4 in Trypanosoma brucei

ABSTRACT: Gene expression in pathogenic protozoans of the family Trypanosomatidae has several novel features, for example in protein synthesis there are multiple eIF4F-like complexes. The eukaryotic eIF4F complex, formed by the eIF4E, eIF4G and eIF4A subunits, is responsible for the canonical selection of mRNAs required for the initiation of mRNA translation. The best-known complexes implicated in translation in trypanosomatids are based on two related pairs of eIF4E and eIF4G subunits (EIF4E3/EIF4G4 and EIF4E4/EIF4G3), whose functional distinctions have yet to be properly identified. Here, to define the interactome associated with both complexes in Trypanosoma brucei procyclic forms, we performed parallel immunoprecipitation experiments followed by identification of proteins co-precipitated with the two sets of tagged eIF4E and eIF4G subunits. A number of different protein partners, including RBPs and RNA helicases, were found to specifically co-precipitate with each complex. Highlights with the EIF4E4/EIF4G3 pair include RBP23, PABP1, EIF4AI and the CRK1 kinase. Co-precipitated partners with the EIF4E3/EIF4G4 pair are more diverse and include DRBD2, PABP2 and different zinc-finger proteins and RNA helicases.EIF4E3/EIF4G4 are known to be essential for viability and to better define their role.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Trypanosoma Brucei

TISSUE(S): Cell Culture

SUBMITTER: MARIA BEZERRA

LAB HEAD: Osvaldo Pompílio de Melo Neto

PROVIDER: PXD033857 | Pride | 2023-03-10

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20170808_59216_Maria_G3IP1_1.RAW	Raw
	20170808_59216_Maria_G3IP1_2.RAW	Raw
	20170808_59216_Maria_G3IP1_3.RAW	Raw
	20170808_59316_Maria_G3IP2_1.RAW	Raw
	20170808_59316_Maria_G3IP2_2.RAW	Raw

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Publications

Distinct mRNA and protein interactomes highlight functional differentiation of major eIF4F-like complexes from <i>Trypanosoma brucei</i>.

Bezerra Maria J R MJR Moura Danielle M N DMN Freire Eden R ER Holetz Fabiola B FB Reis Christian R S CRS Monteiro Tallyta T S TTS Pinto Adriana R S ARS Zhang Ning N Rezende Antonio M AM Pereira-Neves Antonio A Figueiredo Regina C B Q RCBQ Clayton Christine C Field Mark C MC Carrington Mark M de Melo Neto Osvaldo P OP

Frontiers in molecular biosciences 20221007

Gene expression in pathogenic protozoans of the family Trypanosomatidae has several novel features, including multiple eIF4F-like complexes involved in protein synthesis. The eukaryotic eIF4F complex, formed mainly by eIF4E and eIF4G subunits, is responsible for the canonical selection of mRNAs required for the initiation of mRNA translation. The best-known complexes implicated in translation in trypanosomatids are based on two related pairs of eIF4E and eIF4G subunits (EIF4E3/EIF4G4 and EIF4E4/ ...[more]

PMID: 36275617

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Project description:Accumulating data indicate translation plays a role in cancer biology, particularly its rate limiting stage of initiation. Despite this evolving recognition, the function and importance of specific translation initiation factors is unresolved. The eukaryotic translation initiation complex eIF4F consists of eIF4E and eIF4G at a 1:1 ratio. Although it is expected that they display interdependent functions, several publications suggest independent mechanisms. This study is the first to directly assess the relative contribution of eIF4F components to the expressed cellular proteome, transcription factors, microRNAs, and phenotype in a malignancy known for extensive protein synthesis- multiple myeloma (MM). Previously, we have shown that eIF4E/eIF4GI attenuation (siRNA/ Avastin) deleteriously affected MM cells' fate and reduced levels of eIF4E/eIF4GI established targets. Here, we demonstrated that eIF4E/eIF4GI indeed have individual influences on cell proteome. We used an objective, high throughput assay of mRNA microarrays to examine the significance of eIF4E/eIF4GI silencing to several cellular facets such as transcription factors, microRNAs and phenotype. We showed different imprints for eIF4E and eIF4GI in all assayed aspects. These results promote our understanding of the relative contribution and importance of eIF4E and eIF4GI to the malignant phenotype and shed light on their function in eIF4F translation initiation complex. This study concentrated on a particular cancer model and studied the role of eIF4E and eIF4GI in the design of the cells' proteome. We used an unbiased, high throughput system to evaluate the individual importance of eIF4E and eIF4GI levels in MM. We used models of eIF4E or eIF4GI knocked down (KD) MM cell line RPMI 8226 and profiled their respective translated transcription factors (TF), often tumor suppressors or oncogenes. Furthermore, we assessed the KDs' microRNAs repertoires and cells' phenotype. Significant differences were observed between eIF4E and eIF4GI knockdown imprints.