The DIM1 Gene Responsible for the Conserved m62Am62A Dimethylation in the 3′-Terminal Loop of 18 S rRNA is Essential in Yeast

Lafontaine, Denis L. J.; Delcour, Jean; Glasser, Anne-Lise; Desgrès, J; Vandenhaute, Jean

doi:10.1006/jmbi.1994.1525

Cited by 166 publications

(146 citation statements)

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“…However, these phenotypes could be complemented by the corresponding mutated genes, which encoded inactive enzymes (12,59). Likewise, for the dimethylase Dim1p, which catalyzes the formation of the conserved m 2 6 A at positions 1779 and 1780 on yeast 18 S rRNA (60), it was demonstrated that the lethality resulting from deletion of the DIM1 gene was essentially due to a defect in pre-rRNA processing and not to the lack of m 2 6 A at positions 1779 and 1780 on the mature 18 S rRNA (61).…”

Section: Discussionmentioning

confidence: 99%

Lack of Pseudouridine 38/39 in the Anticodon Arm of Yeast Cytoplasmic tRNA Decreases in Vivo Recoding Efficiency

Lecointe¹,

Namy²,

Hatin³

et al. 2002

Journal of Biological Chemistry

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Many different modified nucleotides are found in naturally occurring tRNA, especially in the anticodon region. Their importance for the efficiency of the translational process begins to be well documented. Here we have analyzed the in vivo effect of deleting genes coding for yeast tRNA-modifying enzymes, namely Pus1p, Pus3p, Pus4p, or Trm4p, on termination readthrough and ؉1 frameshift events. To this end, we have transformed each of the yeast deletion strains with a lacZ-luc dual-reporter vector harboring selected programmed recoding sites. We have found that only deletion of the PUS3 gene, encoding the enzyme that introduces pseudouridines at position 38 or 39 in tRNA, has an effect on the efficiency of the translation process. In this mutant, we have observed a reduced readthrough efficiency of each stop codon by natural nonsense suppressor tRNAs. This effect is solely due to the absence of pseudouridine 38 or 39 in tRNA because the inactive mutant protein Pus3[D151A]p did not restore the level of natural readthrough. Our results also show that absence of pseudouridine 39 in the slippery tRNA UAG Leu reduces ؉1 frameshift efficiency. Therefore, the presence of pseudouridine 38 or 39 in the tRNA anticodon arm enhances misreading of certain codons by natural nonsense tRNAs as well as promotes frameshifting on slippery sequences in yeast.

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Section: Discussionmentioning

confidence: 99%

Lack of Pseudouridine 38/39 in the Anticodon Arm of Yeast Cytoplasmic tRNA Decreases in Vivo Recoding Efficiency

Lecointe¹,

Namy²,

Hatin³

et al. 2002

Journal of Biological Chemistry

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“…Dimlp is hghly homologous to KsgAp and can restore both 16s rRNA dimethylation and kasugamycin sensitivity in E. coli (Lafontaine et al 1994). This made it probable that Dimlp is the enzyme responsible for making the homologous dimethylation, m~A,779m~Al,,o, in the yeast 18s rRNA.…”

Section: Dim1 Is Involved In Pre-rrna Processingmentioning

confidence: 99%

“…Association of Dimlp with the pre-ribosomal particle might be required for processing; Dimlp might directly interact with components required for processing or be required for correct ribosome assembly. In either case, this could be mediated through the lysine-rich amino-terminal extension of Dimlp that is not present in the E. coli KsgAp (Lafontaine et al 1994). An alternative possibility is that some factor could have a binding site that overlaps with that of Dimlp.…”

Section: Dim1 Is Involved In Pre-rrna Processingmentioning

confidence: 99%

“…Construction of a conditional GAL10: :dim1 allele A PvuII-XhoI DNA fragment containing the DIMl ORF, 5'-and 3'-flanking sequences (Lafontaine et al 1994) was subcloned into pUC18 to yield plasmid pDL526. In plasmid pDL503, the GAL1-10 promoter region of plasmid pBM272 (Johnston and Davis 1984; kindly provided by M. Johnston, Washington University School of Medicine, St. Louis, MO), isolated by EcoRIBamHI digestion, was subcloned into plasmid pFL44S (Ronneaud et al 1991) in order to be fused to a URA3 marker.…”

Section: Strains and Mediamentioning

confidence: 99%

“…The putative yeast 18s rRNA dimethylase gene, DIMI, was cloned by complementation of the kasugamycin sensitivity of the E. coli ksgA mutant. Dimlp is clearly homologous to KsgAp and can dimethylate the E. coli 1GS rRNA in vivo (Lafontaine et al 1994). In contrast to E. coli ksgA, yeast DIMl is essential for viability, and we therefore generated a conditional allele to determine whether Dimlp is required for ribosome synthesis or function.…”

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confidence: 99%

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The 18S rRNA dimethylase Dim1p is required for pre-ribosomal RNA processing in yeast.

Lafontaine

Vandenhaute²,

Tollervey³

1995

Genes Dev.

187

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The m~A1779m~A178, dimethylation at the 3' end of the small subunit rRNA has been conserved in evolution from bacteria to eukaryotes. The yeast 18s rRNA dimethylase gene DIM1 was cloned previously by complementation in Escherichia coli and shown to be essential for viability in yeast. A conditional GAL1O::diml strain was constructed to allow the depletion of Dimlp from the cell. During depletion, dimethylation of the pre-rRNA is progressively inhibited and pre-rRNA processing at cleavage sites A1 and A2 is concomitantly lost. In consequence, the mature 18s rRNA and its 20s precursor drastically underaccumulate. This has the effect of preventing the synthesis of nonmethylated rRNA. To test whether the processing defect is a consequence of the absence of the dimethylated nucleotides or of the Dimlp dimethylase itself, a cis-acting mutation was created in which both dimethylated adenosines are replaced by guanosine residues. Methylation cannot occur on this mutant pre-rRNA, but no clear pre-rRNA processing defect is seen. Moreover, methylation of the wild-type pre-rRNA predominantly occurs after cleavage at sites A1 and A2. This shows that formation of the m~A , , , , r n~~, ,~, dimethylation is not required for pre-rRNA processing. We propose that the binding of Dimlp to the pre-ribosomal particle is monitored to ensure that only dimethylated pre-rRNA molecules are processed to 18s rRNA.

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Cloning and Characterization of theKlDIM1 Gene fromKluyveromyces lactis Encoding the m26A Dimethylase of the 18S rRNA

Housen

Demonte

Lafontaine

et al. 1997

Yeast

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The DIM1 Gene Responsible for the Conserved m62Am62A Dimethylation in the 3′-Terminal Loop of 18 S rRNA is Essential in Yeast

Cited by 166 publications

References 0 publications

Lack of Pseudouridine 38/39 in the Anticodon Arm of Yeast Cytoplasmic tRNA Decreases in Vivo Recoding Efficiency

Lack of Pseudouridine 38/39 in the Anticodon Arm of Yeast Cytoplasmic tRNA Decreases in Vivo Recoding Efficiency

The 18S rRNA dimethylase Dim1p is required for pre-ribosomal RNA processing in yeast.

Cloning and Characterization of theKlDIM1 Gene fromKluyveromyces lactis Encoding the m26A Dimethylase of the 18S rRNA

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