Domain Structure of tRNase ZL, the Long Form of tRNase Z

tRNase ZL（tRNase Z 的长形式）的结构域结构

• 批准号: 8398288
• 负责人: LOUIS F LEVINGER
• 金额: $ 5.26万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-08 至 2013-08-07
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8398288
• 关键词: Affect; Affinity; Archaea; Bacteria; Binding; Cancer-Predisposing Gene; Catalysis; Cloning; Complement; Complex; Crystallization; Data; Enzymatic Biochemistry; Enzymes; Eukaryota; Evolution; Excision; Holoenzymes; Human; Investigation; Lactamase; Length; Link; Malignant neoplasm of prostate; Mass Spectrum Analysis; Metabolism; Mitochondria; Mutagenesis; Mutation; N Domain; Nuclear; Pathogenesis; Process; Proteins; Proteolysis; Reaction; Resolution; Scheme; Structure; Time; Transfer RNA; Translations; arm; base; dimer; endonuclease; experience; flexibility; monomer; success; tRNA Precursor

DESCRIPTION (provided by applicant): Transfer RNA is central to translation and removal of the 3' trailer is central to tRNA maturation. tRNase Z, the pre-tRNA 3' processing endonuclease, is encoded in a short form (tRNase ZS) in bacteria and archaea and in a long form (tRNase ZL) found only in eukaryotes. As a functioning homodimer, tRNase ZS requires an extensive dimer interface for the subunits to reliably find each other. tRNase ZL originated as a tandem duplication of tRNase ZS which freed the amino and carboxy domains of tRNase ZL to evolve semi-independently, contributing to the ~1,700X greater catalytic efficiency of human tRNase ZL over tRNase ZS. tRNase ZL purifies and presumably functions as a monomer. The amino and carboxy domains of tRNase ZL are linked by a ~70 residue flexible tether. Tethered domains arising from tandem duplications of homodimer proteins may be common and tRNase ZL illustrates the benefits of this evolutionary theme. Flexibility of the tether could also explai the lack of success at crystallizing tRNase ZL. We propose to express and crystallize the amino and carboxy domains of tRNase ZL +/- tether and solve their structures. Success with the domains will guide crystallization of full length tRNase ZL. Complementing domain strategy, formation of a tRNase ZL-tRNA complex could stabilize flexible regions, allowing crystallization of full length tRNase ZL. PUBLIC HEALTH RELEVANCE: tRNase Z has been suggested to be a human prostate cancer susceptibility gene. Numerous pathogenesis-related mutations in human mitochondrial tRNAs affect tRNA metabolism including the tRNase Z reaction. The tandem duplication of tRNase ZS that gave rise to tRNase ZL, including the flexible tether, is an important motif in evolution and enzymology.

描述（由申请人提供）：转移 RNA 对于翻译至关重要，去除 3' 尾部对于 tRNA 成熟至关重要。 tRNase Z 是前 tRNA 3' 加工核酸内切酶，在细菌和古细菌中以短形式 (tRNase ZS) 编码，而长形式 (tRNase ZL) 仅在真核生物中发现。作为一种功能性同二聚体，tRNase ZS 需要广泛的二聚体接口，以便亚基能够可靠地找到彼此。 tRNase ZL 起源于 tRNase ZS 的串联复制，它释放了 tRNase ZL 的氨基和羧基结构域，使其半独立进化，从而使人 tRNase ZL 的催化效率比 tRNase ZS 高约 1,700 倍。 tRNase ZL 纯化并可能作为单体发挥作用。 tRNase ZL 的氨基和羧基结构域通过约 70 个残基的柔性链连接。由同源二聚体蛋白串联重复产生的束缚结构域可能很常见，tRNase ZL 说明了这一进化主题的好处。系链的灵活性也可以解释 tRNase ZL 结晶失败的原因。我们建议表达和结晶 tRNase ZL +/- 系链的氨基和羧基结构域并解析其结构。这些结构域的成功将指导全长 tRNase ZL 的结晶。作为结构域策略的补充，tRNase ZL-tRNA 复合物的形成可以稳定柔性区域，从而允许全长 tRNase ZL 结晶。 公共健康相关性：tRNase Z 已被认为是人类前列腺癌易感基因。人类线粒体 tRNA 中许多与发病机制相关的突变会影响 tRNA 代谢，包括 tRNase Z 反应。 tRNase ZS 的串联复制产生了 tRNase ZL，包括柔性系链，是进化和酶学中的一个重要基序。