Take our Survey

Reference: Urban A, et al. (2009) RNA Sequence and Two-dimensional Structure Features Required for Efficient Substrate Modification by the Saccharomyces cerevisiae RNA:{Psi}-Synthase Pus7p. J Biol Chem 284(9):5845-58

Reference Help

Abstract


The RNA:pseudouridine (psi) synthase Pus7p of Saccharomyces cerevisiae is a multisite-specific enzyme that is able to modify U13 in several yeast tRNAs, U35 in the pre-tRNATyr (GpsiA), U35 in U2 snRNA and U50 in 5S rRNA. Pus7p belongs to the universally conserved TruD-like family of RNA:psi-synthases found in Bacteria, Archaea and Eukarya. While several RNA substrates for yeast Pus7p have been identified, specificity of their recognition and modification has not been studied. However, conservation of an 7 nt-long sequence including the modified U residue in all natural Pus7p substrates suggested importance of these nucleotides for Pus7p recognition and/or catalysis. Using site-directed mutagenesis we designed a set of RNA variants derived from the yeast tRNAAsp(GUC), pre-tRNATyr(GpsiA) and U2 snRNA and tested their ability to be modified by Pus7p in vitro. We demonstrated that the highly conserved U-2 and A+1 residues (as referred to modified U0) are crucial identity elements for efficient modification by Pus7p. Nucleotide substitutions at other surrounding positions (-4, -3, +2, +3) have only a moderate effect. Surprisingly, the identity of the nucleotide immediately 5' to the target U0 residue (position -1) is not important for efficient modification. Alteration of tRNA 3D structure had no detectable effect on Pus7p activity at position 13. However, our results suggest that the presence at least of one stem-loop structure including or close to the target U nucleotide is required for Pus7p-catalyzed modification.

Reference Type
Journal Article
Authors
Urban A, Behm-Ansmant I, Branlant C, Motorin Y
Primary Lit For
Additional Lit For
Review For

Interaction Annotations


Increase the total number of rows showing on this page by using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i" buttons located within a cell for an annotation to view further details about experiment type and any other genes involved in the interaction.

Interactor Interactor Type Assay Annotation Action Modification Phenotype Source Reference

Gene Ontology Annotations


Increase the total number of rows showing on this page using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table.

Gene Gene Ontology Term Qualifier Aspect Method Evidence Source Assigned On Annotation Extension Reference

Phenotype Annotations


Increase the total number of rows showing on this page using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i" buttons located within a cell for an annotation to view further details.

Gene Phenotype Experiment Type Mutant Information Strain Background Chemical Details Reference

Regulation Annotations


Increase the total number of rows displayed on this page using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; to filter the table by a specific experiment type, type a keyword into the Filter box (for example, “microarray”); download this table as a .txt file using the Download button or click Analyze to further view and analyze the list of target genes using GO Term Finder, GO Slim Mapper, SPELL, or YeastMine.

Regulator Target Experiment Assay Construct Conditions Strain Background Reference