Metal content and kinetic properties of yeast RNA lariat debranching enzyme Dbr1.

ABSTRACT: In eukaryotic cells, intron lariats produced by the spliceosome contain a 2'5' phosphodiester linkage. The RNA lariat debranching enzyme, Dbr1, is the only enzyme known to hydrolyze this bond. Dbr1 is a member of the metallophosphoesterase (MPE) family of enzymes, and recent X-ray crystal structures and biochemistry data demonstrate that Dbr1 from Entamoeba histolytica uses combinations of Mn²⁺, Zn²⁺, and Fe²⁺ as enzymatic cofactors. Here, we examine the kinetic properties and metal dependence of the Dbr1 homolog from Saccharomyces cerevisiae (yDbr1). Elemental analysis measured stoichiometric quantities of Fe and Zn in yDbr1 purified following heterologous expression E. coli We analyzed the ability of Fe²⁺, Zn²⁺,

SUBMITTER: Clark NE 

PROVIDER: S-EPMC9202583 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature