3P1Z

Crystal structure of the Aperopyrum pernix RNA splicing endonuclease

3P1Z の概要

• エントリーDOI: 10.2210/pdb3p1z/pdb
• 関連するPDBエントリー: 3P1Y
• 分子名称: Putative uncharacterized protein, tRNA-splicing endonuclease (3 entities in total)
• 機能のキーワード: mixed antiparallel and parallel beta-sheet, heterotetramer, rna splicing, rna, splicing endonuclease, hydrolase
• 由来する生物種: Aeropyrum pernix; 詳細
• タンパク質・核酸の鎖数: 12
• 化学式量合計: 235368.05

構造登録者

Hirata, A. (登録日: 2010-10-01, 公開日: 2011-08-10, 最終更新日: 2024-10-30)

主引用文献

Hirata, A.,Kitajima, T.,Hori, H.
Cleavage of intron from the standard or non-standard position of the precursor tRNA by the splicing endonuclease of Aeropyrum pernix, a hyper-thermophilic Crenarchaeon, involves a novel RNA recognition site in the Crenarchaea specific loop
Nucleic Acids Res., 39:9376-9389, 2011

PubMed Abstract: In Crenarchaea, several tRNA genes are predicted to express precursor-tRNAs (pre-tRNAs) with canonical or non-canonical introns at various positions. We initially focused on the tRNA(Thr) species of hyperthermophilic crenarchaeon, Aeropyrum pernix (APE) and found that in the living APE cells three tRNA(Thr) species were transcribed and subsequently matured to functional tRNAs. During maturation, introns in two of them were cleaved from standard and non-standard positions. Biochemical studies revealed that the APE splicing endonuclease (APE-EndA) removed both types of introns, including the non-canonical introns, without any nucleotide modification. To clarify the underlying reasons for broad substrate specificity of APE-EndA, we determined the crystal structure of wild-type APE-EndA and subsequently compared its structure with that of Archaeaoglobus fulgidus (AFU)-EndA, which has narrow substrate specificity. Remarkably, structural comparison revealed that APE-EndA possesses a Crenarchaea specific loop (CSL). Introduction of CSL into AFU-EndA enhanced its intron-cleaving activity irrespective of the position or motif of the intron. Thus, our biochemical and crystallographic analyses of the chimera-EndA demonstrated that the CSL is responsible for the broad substrate specificity of APE-EndA. Furthermore, mutagenesis studies revealed that Lys44 in CSL functions as the RNA recognition site.

PubMed: 21846775
DOI: 10.1093/nar/gkr615

実験手法

X-RAY DIFFRACTION (2.8 Å)