A novel three-unit tRNA splicing endonuclease found in ultrasmall Archaea possesses broad substrate specificity

作者：Fujishima Kosuke; Sugahara Junichi; Miller Christopher S; Baker Brett J; Di Giulio Massimo; Takesue Kanako; Sato Asako; Tomita Masaru; Banfield Jillian F; Kanai Akio^*

来源：Nucleic Acids Research, 2011, 39(22): 9695-9704.

DOI：10.1093/nar/gkr692

tRNA splicing endonucleases, essential enzymes found in Archaea and Eukaryotes, are involved in the processing of pre-tRNA molecules. In Archaea, three types of splicing endonuclease [homotetrameric: alpha(4), homodimeric: alpha(2), and heterotetrameric: (alpha beta)(2)] have been identified, each representing different substrate specificity during the tRNA intron cleavage. Here, we discovered a fourth type of archaeal tRNA splicing endonuclease (epsilon(2)) in the genome of the acidophilic archaeon Candidatus Micrarchaeum acidiphilum, referred to as ARMAN-2 and its closely related species, ARMAN-1. The enzyme consists of two duplicated catalytic units and one structural unit encoded on a single gene, representing a novel three-unit architecture. Homodimeric formation was confirmed by cross-linking assay, and site-directed mutagenesis determined that the conserved L10-pocket interaction between catalytic and structural unit is necessary for the assembly. A tRNA splicing assay reveal that epsilon(2) endonuclease cleaves both canonical and non-canonical bulge-helix-bulge motifs, similar to that of (alpha beta)(2) endonuclease. Unlike other ARMAN and Euryarchaeota, tRNAs found in ARMAN-2 are highly disrupted by introns at various positions, which again resemble the properties of archaeal species with (alpha beta)(2) endonuclease. Thus, the discovery of epsilon(2) endonuclease in an archaeon deeply branched within Euryarchaeota represents a new example of the coevolution of tRNA and their processing enzymes.

• 出版日期2011-12