Repressor binding to a dorsal regulatory site traps human eIF4E in a high cap-affinity state

Ptushkina, Marina; Haar, Tobias von der; Karim, Muhammad Manjurul; Hughes, Juliette H.; McCarthy, John E.G.

doi:10.1093/emboj/18.14.4068

Cited by 123 publications

(138 citation statements)

References 32 publications

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“…For both eIF4E preparations, a binding constant of 1.1 ϫ 10 Ϫ8 M was determined using the Biacore evaluation software to fit the three curves (global fit, 1:1 Langmuir binding). These values are in very good agreement with those reported by Ptushkina et al (19) (1.0 Ϯ 0.4 ϫ 10 Ϫ8 ). Thus, phosphorylation of eIF4E does not appear to influence its ability to interact with 4E-BP1.…”

Section: Gtp Interactionsupporting

confidence: 93%

“…Association of eIF4E with 4E-BP1 or eIF4G has been shown to stabilize its interaction with capped mRNA (19,21). Consistent with this, association of eIF4E with 4E-BP1 reduced its rate of dissociation from capped RNA by 70-fold (Table II).…”

Section: E-bp1 Stabilizes the Binding Of Phosphorylated Eif4e Tomentioning

confidence: 53%

“…Thus, phosphorylation of eIF4E does not appear to influence its ability to interact with 4E-BP1. Since the same region of eIF4E is involved in its interactions with 4E-BP1 and eIF4G (1,19), it is highly likely that phosphorylation of eIF4E also does not affect binding to eIF4G, although we have been unable to test this directly.…”

Section: Gtp Interactionmentioning

confidence: 93%

“…Biotinylated capped and uncapped RNAs were prepared by in vitro transcription in the presence or absence of m 7 GpppG, respectively, with the same template as described previously (19), which was kindly provided by Dr. Tobias von der Haar (University of Manchester Institute of Science and Technology, Manchester, UK). The RNAs contain only one uridine (in the 3Ј-end), allowing synthesis of singly 3Ј-biotinylated transcripts.…”

Section: Methodsmentioning

confidence: 99%

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Phosphorylation of Eukaryotic Initiation Factor 4E Markedly Reduces Its Affinity for Capped mRNA

Scheper¹,

Kollenburg²,

Hu³

et al. 2002

Journal of Biological Chemistry

230

198

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In eukaryotes, a key step in the initiation of translation is the binding of the eukaryotic initiation factor 4E (eIF4E) to the cap structure of the mRNA. Subsequent recruitment of several components, including the small ribosomal subunit, is thought to allow migration of initiation complexes and recognition of the initiation codon. Mitogens and cytokines stimulate the phosphorylation of eIF4E at Ser 209 , but the functional consequences of this modification have remained a major unresolved question. Using fluorescence spectroscopy and surface plasmon resonance techniques, we show that phosphorylation of eIF4E markedly reduces its affinity for capped RNA, primarily due to an increased rate of dissociation. Variant eIF4E proteins harboring negatively charged acidic residues at position 209 also showed decreased binding to capped RNA. Furthermore, a basic residue at position 159 was shown to be essential for cap binding. Although eIF4E-binding protein 1 greatly stabilized binding of phosphorylated eIF4E to capped RNA, in the presence of eIF4E-binding protein 1 the phosphorylated form still dissociated faster compared with nonphopshorylated eIF4E. The implications of our findings for the mechanism of translation initiation are discussed.In eukaryotic cells, all nucleus-encoded mRNAs possess a so-called "cap structure" at their 5Ј-end. This cap consists of a methylated 5Ј-5Ј bound guanosine triphosphate (m 7 GTP) moiety. The cap plays a key role in the translation of the mRNA by permitting recruitment of the eukaryotic initiation factors (eIFs) 1 required for the attachment of the ribosome and correct initiation of translation. The protein that interacts directly with the cap is eIF4E, which forms a complex with the scaffolding protein eIF4G, which in turn recruits other factors. These include eIF4A and eIF4B, which are involved in unwinding secondary structure in the 5Ј-untranslated region of the mRNA, and the poly(A)-binding protein, which by interacting with the 3Ј-tail of the mRNA circularizes it (1, 2). eIF4E also binds to small regulatory proteins termed eIF4E-binding proteins (4E-BPs). These compete with eIF4G for binding to overlapping sites on eIF4E and thus inhibit formation of initiation complexes (3, 4).It has long been known that eIF4E undergoes regulated phosphorylation, and the site has been identified as Ser 209 (5, 6). Phosphorylation of eIF4E is increased by mitogenic stimuli that activate translation (reviewed in Ref. 7) and by cytokines (8, 9). Recently, two kinases were identified that phosphorylate Ser 209 in eIF4E and are targets for the mitogen-activated extracellular signal-regulated kinase and stress/cytokine-activated p38 mitogen-activated protein kinase pathways (9 -13). These enzymes (Mnk1 and Mnk2) also associate with eIF4G in vivo (11,12,14). Despite these advances and the large number of studies on the changes of eIF4E phosphorylation in vivo, the key issue of the effect of phosphorylation on the function of eIF4E has received little attention and has remained a major question in the ...

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Section: Gtp Interactionsupporting

confidence: 93%

Section: E-bp1 Stabilizes the Binding Of Phosphorylated Eif4e Tomentioning

confidence: 53%

Section: Gtp Interactionmentioning

confidence: 93%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Phosphorylation of Eukaryotic Initiation Factor 4E Markedly Reduces Its Affinity for Capped mRNA

Scheper¹,

Kollenburg²,

Hu³

et al. 2002

Journal of Biological Chemistry

230

198

View full text Add to dashboard Cite

show abstract

“…Thus, altering the Y76-W130 interaction could be the initial step in a chain of events communicating Z binding to the cap-binding site. Interestingly, W130 was proposed to be part of an allosteric track important for communication between the dorsal surface and the cap-binding site (28). Given the differing binding surfaces for Z and eIF4Gp, communication to the cap-binding site may go through different allosteric routes and thereby yield different outcomes.…”

Section: Resultsmentioning

confidence: 99%

Structural characterization of the Z RING-eIF4E complex reveals a distinct mode of control for eIF4E

Volpon

Osborne

Capul

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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The eukaryotic translation initiation factor eIF4E, a potent oncogene, is highly regulated. One class of eIF4E regulators, including eIF4G and the 4E-binding proteins (4E-BPs), interact with eIF4E using a conserved YXXXXLΦ-binding site. The structural basis of this interaction and its regulation are well established. Really Interesting New Gene (RING) domain containing proteins, such as the promyelocytic leukemia protein PML and the arenaviral protein Z, represent a second class of eIF4E regulators that inhibit eIF4E function by decreasing eIF4E's affinity for its m 7 G cap ligand. To elucidate the structural basis of this inhibition, we determined the structure of Z and studied the Z-eIF4E complex using NMR methods. We show that Z interacts with eIF4E via a novel binding site, which has no homology with that of eIF4G or the 4E-BPs, and is different from the RING recognition site used in the ubiquitin system. Z and eIF4G interact with distinct parts of eIF4E and differentially alter the conformation of the m 7 G cap-binding site. Our results provide a molecular basis for how PML and Z RINGs reduce the affinity of eIF4E for the m 7 G cap and thereby act as key inhibitors of eIF4E function. Furthermore, our findings provide unique insights into RING protein interactions.Lassa Fever Virus-Z | NMR | promyelocytic leukemia protein

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Survey of the 1999 surface plasmon resonance biosensor literature

2000

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The application of surface plasmon resonance biosensors in life sciences and pharmaceutical research continues to increase. This review provides a comprehensive list of the commercial 1999 SPR biosensor literature and highlights emerging applications that are of general interest to users of the technology. Given the variability in the quality of published biosensor data, we present some general guidelines to help increase confidence in the results reported from biosensor analyses. Copyright © 2000 John Wiley & Sons, Ltd.

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Repressor binding to a dorsal regulatory site traps human eIF4E in a high cap-affinity state

Cited by 123 publications

References 32 publications

Phosphorylation of Eukaryotic Initiation Factor 4E Markedly Reduces Its Affinity for Capped mRNA

Phosphorylation of Eukaryotic Initiation Factor 4E Markedly Reduces Its Affinity for Capped mRNA

Structural characterization of the Z RING-eIF4E complex reveals a distinct mode of control for eIF4E

Survey of the 1999 surface plasmon resonance biosensor literature

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