Mechanism of Group II Intron Retrotransposition

II组内含子逆转录转座机制

• 批准号: 6741190
• 负责人: LORI M CONLAN
• 金额: $ 4.3万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6741190
• 关键词: DNA replication; RNA directed DNA polymerase; Streptococcus lactis; electron crystallography; endonuclease; genetic transcription; introns; microarray technology; nucleic acid repetitive sequence; postdoctoral investigator; protein structure function; reverse transcriptase inhibitors; transposon /insertion element

DESCRIPTION (provided by applicant): Mobile group II introns are retroelements that are widely hypothesized to be related to both non-LTR retrotransposons and spliceosomal introns. The overall goal of this project is to combine genetic, biochemical and structural studies to characterize the Lactococcus lactis group II intron-encoded protein (LtrA) and the pathways involved in intron mobility, particularly retrotransposition. To accomplish this, there are two specific aims. First, detailed biochemical analyses on the structural domains of LtrA will be performed. The relationship of the maturase, endonuclease, and reverse transcriptase activities to domains will be explored to understand the modular composition and evolution of LtrA. Isolated domains will also prove useful for crystallography. Second, to better understand the mechanisms of intron retrotransposition, a selectable genetic assay for isolated retrotransposition events of the group II L. lactis intron LtrB will be used. Different plasmid recipients will be employed to explore the nucleic acid targets of intron transposition and the dependency on the processes of replication and/or transcription. I will therefore understand the basic principles of group II intron biology and their relationship, as appropriate, to non-LTR retrotransposons and spliceosomal introns.

描述（由申请人提供）：移动的II组内含子是广泛假设与非LTR反转录转座子和剪接体内含子相关的反转录元件。该项目的总体目标是联合收割机的遗传，生物化学和结构的研究，以表征乳酸乳球菌组II内含子编码蛋白（LtrA）和参与内含子移动，特别是反转录转座的途径。为了实现这一点，有两个具体目标。首先，将对LtrA的结构域进行详细的生化分析。成熟酶，核酸内切酶和逆转录酶活性域的关系将被探讨，以了解模块组成和进化的LtrA。孤立的域也将被证明是有用的结晶学。第二，为了更好地了解内含子反转录转座的机制，对II L组的孤立反转录转座事件进行了选择性遗传分析。将使用lactis内含子LtrB。将采用不同的质粒受体来探索内含子转座的核酸靶点以及对复制和/或转录过程的依赖性。因此，我将了解第二组内含子生物学的基本原则，以及它们与非LTR反转录转座子和剪接体内含子的关系。