Structural basis for LeishIF4E-1 modulation by an interacting protein in the human parasite Leishmania major

Meleppattu, Shimi; Arthanari, Haribabu; Zinoviev, Alexandra; Boeszoermenyi, Andras; Wagner, Gerhard; Shapira, Michael Y.; Léger-Abraham, Mélissa

doi:10.1093/nar/gky194

Cited by 19 publications

(34 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…An alternative is that EIF4E1-4EIP complex is functionally similar to 4EHP-GYF2 and that EIF4E1 acts as a constitutively inhibitory translation factor ( Fig 9B), with mRNA-binding enhanced by cooperative interactions with 4EIP. If the 4EIP interaction prevents cap binding by EIF4E1 (65), this might allow decapping ( Fig 9C).…”

Section: Discussionmentioning

confidence: 99%

“…Further, it was hypothesised that the initiation activity of EIF4E1 is suppressed, in the promastigote stage, by interaction with 4EIP (63). The results of yeast two-hybrid assays indicated that a YXXXXLØ motif at the N-terminus of Leishmania 4EIP was required for interaction with EIF4E1 (63), and co-crystallization revealed that first 52 residues of 4EIP form two alpha-helices that interact with EIF4E1; the first includes the consensus motif (65). In vitro evidence suggested that binding of m 7 GTP by EIF4E1 was inhibited by addition of the N-terminal 4EIP fragment, probably as a consequence of a conformational change within the cap-binding pocket (65).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The suppressive cap-binding-complex factor 4EIP is required for normal differentiation

Clayton

Terrao

Marucha

et al. 2018

Preprint

View full text Add to dashboard Cite

Summary/AbstractTrypanosoma brucei live in mammals as bloodstream forms and in the Tsetse midgut as procyclic forms. Differentiation from one form to the other proceeds via a growth-arrested stumpy form with low mRNA content and translation. The parasites have six eIF4Es and five eIF4Gs. EIF4E1 pairs with the mRNA-binding protein 4EIP but not with any EIF4G. EIF4E1 and 4EIP each inhibit expression when tethered to a reporter mRNA. The 4E-binding motif in 4EIP is required for the interaction with EIF4E1 both in vivo and in a 2-hybrid assay, but not for the suppressive activity of 4EIP when tethered. However, the suppressive activity of EIF4E1 when tethered requires 4EIP. Correspondingly, in growing bloodstream forms, 4EIP is preferentially associated with unstable mRNAs. Trypanosomes lacking 4EIP have a marginal growth disadvantage as cultured bloodstream or procyclic forms. Bloodstream forms without 4EIP cannot make differentiation-competent stumpy forms, but the defect can be complemented by a truncated 4EIP that does not interact with EIF4E1. Bloodstream forms lacking EIF4E1 have a growth defect but can differentiate. We suggest that 4EIP and EIF4E1 fine-tune mRNA levels in growing cells, and that 4EIP is required for mRNA suppression during differentiation to the stumpy form.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The suppressive cap-binding-complex factor 4EIP is required for normal differentiation

Clayton

Terrao

Marucha

et al. 2018

Preprint

View full text Add to dashboard Cite

show abstract

“…The 3-D structure of LeishIF4E1 was recently solved, when it was associated with a fragment derived from its unique binding partner Leish4E-IP1. Semi- in vivo binding assays suggested that recombinant Leish4E-IP1 inhibits the cap-binding activity of LeishIF4E1 [16], suggesting that 4E-IP1 is a functional inhibitor of LeishIF4E1.…”

Section: Introductionmentioning

confidence: 99%

Distinct features of theLeishmaniacap-binding protein LeishIF4E2 revealed by CRISPR-Cas9 mediated heterozygous deletion

Baron

Tupperwar

Davidov

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

20 Leishmania parasites cycle between sand-fly vectors and mammalian hosts, adapting to 21 alternating environments by stage-differentiation, accompanied by changes in the proteome 22 profiles. Translation regulation plays a central role in driving the differential program of gene 23 expression, since control of gene regulation in Leishmania is mostly post-transcriptional. 24 The Leishmania genome encodes six eIF4E candidates, each assumed to have specific functions, 25 although overlaps are expected. It is noted that some of them can bind to a dedicated eIF4G 26 candidate partners, and LeishIF4E2 does not bind any known eIF4G ortholog. LeishIF4E2 was 27 previously shown to comigrate in the polysomal fractions of sucrose gradients, whereas initiation 28 factors usually comigrate with pre-initiation and initiation complexes. Using the CRISPR-Cas9 29 methodology, we deleted one of the two LeishIF4E2 gene copies. The deletion caused severe 30 alterations in the morphology of mutant cells that turned round and equipped with a very short 31 flagellum, but their growth rate and general translation remained unaffected. Proteomic analysis 32 of the LeishIF4E2(+/-) mutant cells compared to wild type controls showed that the number of 33 proteins that were upregulated exceeded the number of downregulated proteins, possibly 34 indicating that a repressor function was eliminated. The upregulated proteins were related mainly 35 to general metabolic processes, DNA repair and replication, signaling, and cellular motor 36 activity. The downregulated proteins included several groups, including cytoskeletal and 37 ribosomal proteins. Despite the fact that only one of the two LeishIF4E2 alleles was deleted, the 38 mutant cells were impaired in their ability to infect cultured macrophages. LeishIF4E2 does not 39 behave like a general translation factor and its function remains elusive. Although it could have a 40 repressive function, we cannot exclude the possibility that it is responsible for translation of a 41 specific set of transcripts. Overall, our results are in line with the possibility that the different 42 LeishIF4Es are assigned unique functions. 43 44 Author summary 45 Leishmania parasites cause a broad spectrum of diseases that lead to different 46 pathological symptoms. During their life cycle, the parasites shuffle between sand-fly 47 vectors and mammalian hosts, while adapting to changing environments via a stage 48 specific program of gene expression that assists the survival of Leishmania under the 49 changing conditions. Translation initiation plays a key role in control of gene expression, 50 in Leishmania this is exemplified by the presence of multiple cap-binding complexes that 51 interact with mRNAs. The parasites encode multiple paralogs of the cap-binding 52 translation initiation factor eIF4E, and of its corresponding binding partner eIF4G, 53 forming complexes with different potential functions. Using the CRISPR-Cas9 54 methodology, we generated a heterozygous mutant of the least studied cap-binding 55 pa...

show abstract

“…Additionally, 4E-IP which is constitutively expressed, only interacts with eIF4E1 during the promastigote stage [90]. It has been proposed that 4E-IP allosterically destabilizes interaction of Leishmania eIF4E1 with the cap4 structure [91]. Bloodstream forms of the parasite lacking 4E-IP are defective in translational suppression and cannot develop into the procyclic form which initiates the promastigote life cycle [98].…”

Section: Eif4e and Interactors From Protozoan Parasitesmentioning

confidence: 99%

eIF4E and Interactors from Unicellular Eukaryotes

Ross‐Kaschitza

Altmann

2020

IJMS

View full text Add to dashboard Cite

eIF4E, the mRNA cap-binding protein, is well known as a general initiation factor allowing for mRNA-ribosome interaction and cap-dependent translation in eukaryotic cells. In this review we focus on eIF4E and its interactors in unicellular organisms such as yeasts and protozoan eukaryotes. In a first part, we describe eIF4Es from yeast species such as Saccharomyces cerevisiae, Candida albicans, and Schizosaccharomyces pombe. In the second part, we will address eIF4E and interactors from parasite unicellular species—trypanosomatids and marine microorganisms—dinoflagellates. We propose that different strategies have evolved during evolution to accommodate cap-dependent translation to differing requirements. These evolutive “adjustments” involve various forms of eIF4E that are not encountered in all microorganismic species. In yeasts, eIF4E interactors, particularly p20 and Eap1 are found exclusively in Saccharomycotina species such as S. cerevisiae and C. albicans. For protozoan parasites of the Trypanosomatidae family beside a unique cap4-structure located at the 5′UTR of all mRNAs, different eIF4Es and eIF4Gs are active depending on the life cycle stage of the parasite. Additionally, an eIF4E-interacting protein has been identified in Leishmania major which is important for switching from promastigote to amastigote stages. For dinoflagellates, little is known about the structure and function of the multiple and diverse eIF4Es that have been identified thanks to widespread sequencing in recent years.

show abstract

Structural basis for LeishIF4E-1 modulation by an interacting protein in the human parasite Leishmania major

Cited by 19 publications

References 59 publications

The suppressive cap-binding-complex factor 4EIP is required for normal differentiation

The suppressive cap-binding-complex factor 4EIP is required for normal differentiation

Distinct features of theLeishmaniacap-binding protein LeishIF4E2 revealed by CRISPR-Cas9 mediated heterozygous deletion

eIF4E and Interactors from Unicellular Eukaryotes

Contact Info

Product

Resources

About