Dissecting the Ribosomal Inhibition Mechanisms of Edeine and Pactamycin

Dinos, George P.; Wilson, Daniel N.; Teraoka, Yoshika; Szaflarski, Witold; Fucini, Paola; Kalpaxis, Dimitrios L.; Nierhaus, Knud H.

doi:10.1016/s1097-2765(04)00002-4

Cited by 147 publications

(74 citation statements)

References 32 publications

(2 reference statements)

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“…7E and F). Similarly, pactamycin, which prevents polysome formation by blocking translocation (36), led to a shift of all three investigated RNAs into the light fractions of the gradient (Fig. 7H and I).…”

Section: Resultsmentioning

confidence: 76%

CstF64: Cell Cycle Regulation and Functional Role in 3′ End Processing of Replication-Dependent Histone mRNAs

Romeo

Griesbach

Schümperli

2014

Molecular and Cellular Biology

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The 3= end processing of animal replication-dependent histone mRNAs is activated during G 1 /S-phase transition. The processing site is recognized by stem-loop binding protein and the U7 snRNP, but cleavage additionally requires a heat-labile factor (HLF), composed of cleavage/polyadenylation specificity factor, symplekin, and cleavage stimulation factor 64 (CstF64). Although HLF has been shown to be cell cycle regulated, the mechanism of this regulation is unknown. Here we show that levels of CstF64 increase toward the S phase and its depletion affects histone RNA processing, S-phase progression, and cell proliferation. Moreover, analyses of the interactions between CstF64, symplekin, and the U7 snRNP-associated proteins FLASH and Lsm11 indicate that CstF64 is important for recruiting HLF to histone precursor mRNA (pre-mRNA)-resident proteins. Thus, CstF64 is central to the function of HLF and appears to be at least partly responsible for its cell cycle regulation. Additionally, we show that misprocessed histone transcripts generated upon CstF64 depletion mainly accumulate in the nucleus, where they are targets of the exosome machinery, while a small cytoplasmic fraction is partly associated with polysomes.

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Section: Resultsmentioning

confidence: 76%

CstF64: Cell Cycle Regulation and Functional Role in 3′ End Processing of Replication-Dependent Histone mRNAs

Romeo

Griesbach

Schümperli

2014

Molecular and Cellular Biology

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“…In contrast, the HCV IRES recruits the 40S ribosomal subunit directly to the initiation codon without scanning. Therefore, we used edeine, a strongly basic, linear oligopeptide antibiotic that interferes with initiation codon (AUG) recognition by the scanning 40S-ternary complexes (24,25,62,63), to determine whether the 40S ribosomal subunit was recruited by the HTLV-1 IRES directly at the initiation codon or upstream of it. At low concentrations, edeine inhibits translation initiation mediated by scanning 40S ribosomal subunits in RRL, with no effect on polypeptide elongation (25,64).…”

Section: Resultsmentioning

confidence: 99%

“…By using the drug edeine, which inhibits the initiation step of protein synthesis by blocking the recognition of the initiation codon by scanning ribosomes (23,24,62,66), we provide insight into the strategy used by the HTLV-1 IRES to recruit the initiation complex (Fig. 13).…”

Section: Discussionmentioning

confidence: 99%

“…In addition, evidence is provided demonstrating that the HTLV-1 IRES is dependent on ribosomal protein S25 for full activity. The HTLV-1 IRES is sensitive to edeine, a drug that inhibits the initiation step of protein synthesis by impeding the recognition of the initiation codon by scanning 40S ribosomal subunits (23)(24)(25), suggesting that upon internal recruitment, the initiation complex follows a scanning mechanism leading to the recognition of the initiation codon of the HTLV-1 Gag open reading frame.…”

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confidence: 99%

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The 5′ Untranslated Region of the Human T-Cell Lymphotropic Virus Type 1 mRNA Enables Cap-Independent Translation Initiation

Olivares

Landry

Cáceres

et al. 2014

J Virol

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The human T-cell leukemia virus type 1 (HTLV-1) is a complex human retrovirus that causes adult T cell leukemia and of HTLVassociated myelopathy/tropical spastic paraparesis. The mRNA of some complex retroviruses, including the human and simian immunodeficiency viruses (HIV and SIV), can initiate translation using a canonical cap-dependent mechanism or through an internal ribosome entry site (IRES). In this study, we present strong evidence showing that like HIV-1 and SIV, the 5=-untranslated region (5=UTR) of the HTLV-1 full-length mRNA harbors an IRES. Cap-independent translational activity was evaluated and demonstrated using dual luciferase bicistronic mRNAs in rabbit reticulocyte lysate, in mammalian cell culture, and in Xenopus laevis oocytes. Characterization of the HTLV-1 IRES shows that its activity is dependent on the ribosomal protein S25 (RPS25) and that its function is highly sensitive to the drug edeine. Together, these findings suggest that the 5=UTR of the HTLV-1 full-length mRNA enables internal recruitment of the eukaryotic translation initiation complex. However, the recognition of the initiation codon requires ribosome scanning. These results suggest that, after internal recruitment by the HTLV-1 IRES, a scanning step takes place for the 40S ribosomal subunit to be positioned at the translation initiation codon. IMPORTANCEThe mechanism by which retroviral mRNAs recruit the 40S ribosomal subunit internally is not understood. This study provides new insights into the mechanism of translation initiation used by the human T-cell lymphotropic virus type 1 (HTLV-1). The results show that the HTLV-1 mRNA can initiate translation via a noncanonical mechanism mediated by an internal ribosome entry site (IRES). This study also provides evidence showing the involvement of cellular proteins in HTLV-1 IRES-mediated translation initiation. Together, the data presented in this report significantly contribute to the understanding of HTLV-1 gene expression.

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“…In the small ribosomal subunit, antibiotics act upon a variety of sites where the drugs may affect binding of tRNAs, displace mRNA, or interfere with translocation (11)(12)(13)(14)(15)(16). In a striking contrast, the distribution of sites of antibiotic action in the large ribosomal subunit is highly constrained.…”

mentioning

confidence: 99%

Potential New Antibiotic Sites in the Ribosome Revealed by Deleterious Mutations in RNA of the Large Ribosomal Subunit

Yassin¹,

Mankin

2007

Journal of Biological Chemistry

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The ribosome is the main target for antibiotics that inhibit protein biosynthesis. Despite the chemical diversity of the known antibiotics that affect functions of the large ribosomal subunit, these drugs act on only a few sites corresponding to some of the known functional centers. We have used a genetic approach for identifying structurally and functionally critical sites in the ribosome that can be used as new antibiotic targets. By using randomly mutagenized rRNA genes, we mapped rRNA sites where nucleotide alterations impair the ribosome function or assembly and lead to a deleterious phenotype. A total of 77 single-point deleterious mutations were mapped in 23 S rRNA and ranked according to the severity of their deleterious phenotypes. Many of the mutations mapped to familiar functional sites that are targeted by known antibiotics. However, a number of mutations were located in previously unexplored regions. The distribution of the mutations in the spatial structure of the ribosome showed a strong bias, with the strongly deleterious mutations being mainly localized at the interface of the large subunit and the mild ones on the solvent side. Five sites where deleterious mutations tend to cluster within discrete rRNA elements were identified as potential new antibiotic targets. One of the sites, the conserved segment of helix 38, was studied in more detail. Although the ability of the mutant 50 S subunits to associate with 30 S subunits was impaired, the lethal effect of mutations in this rRNA element was unrelated to its function as an intersubunit bridge. Instead, mutations in this region had a profound deleterious effect on the ribosome assembly.

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Dissecting the Ribosomal Inhibition Mechanisms of Edeine and Pactamycin

Cited by 147 publications

References 32 publications

CstF64: Cell Cycle Regulation and Functional Role in 3′ End Processing of Replication-Dependent Histone mRNAs

CstF64: Cell Cycle Regulation and Functional Role in 3′ End Processing of Replication-Dependent Histone mRNAs

The 5′ Untranslated Region of the Human T-Cell Lymphotropic Virus Type 1 mRNA Enables Cap-Independent Translation Initiation

Potential New Antibiotic Sites in the Ribosome Revealed by Deleterious Mutations in RNA of the Large Ribosomal Subunit

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