GENETIC AND MOLECULAR STUDIES OF GROUP II INTRONS

II 组内含子的遗传和分子研究

• 批准号: 6924216
• 负责人: RONALD A BUTOW
• 金额: $ 21.06万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-08-01 至 2005-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6924216
• 关键词: RNA binding protein; RNA directed DNA polymerase; RNA splicing; Saccharomyces; complementary DNA; endonuclease; fungal genetics; gene mutation; genetic recombination; introns; laboratory rabbit; mitochondria; mitochondrial DNA; monoclonal antibody; nucleic acid sequence; open reading frames; precursor mRNA; protein structure function

Splicing and genetic properties of group II introns will be investigated in mitochondria of bakers yeast, Saccharomyces cerevisiae. Introns I and 2 of the COXI gene of mtDNA code for a protein with reverse transcriptase (RT), endonuclease and maturase (splicing) functions essential for the splicing and homing/transposition by the introns. Both introns are highly efficient, site-specific retroelements clearly related to non-LTR- retrotransposons, including the LINE elements of the human genome. Major and minor intron homing pathways have been identified, all of which depend on a remarkable reaction in which a complex containing the intron-encoded RT and excised intron RNA lariat inserts the intron RNA into the sense strand of the intron-less DNA target site by a reverse splicing reaction. Subsequent steps vary widely among various RT- independent pathways. Bakers yeast has numerous attributes that make it a powerful and facile system for research on mechanisms of group II intron homing, and for the mechanisms of site-specific homing by group II introns to obtain a clear view of how the various pathways differ and, or importantly, to show how a common homing intermediate is partitioned among processes, transposition was likely a mechanism by which introns spread during evolution. A system will be developed to detect retrotransposition of a tagged intron from mitochondria to the nucleus. The third aim is to characterize aspects of group II intron molecular biology. The main emphasis is to identify nuclear will analyze genes that process the precursor to the intron-encoded RT, control the stability and degradation of excised group II intron RNAs, and assist the self-splicing of the introns. Finally, the RNA binding and splicing functions of the naturase domain of the intron-encoded protein will be studied.

第二组内含子的剪接和遗传特性将在面包酵母，酿酒酵母的线粒体中进行研究。线粒体COXI基因的内含子I和2编码具有逆转录酶（RT）、核酸内切酶和成熟酶（剪接）功能的蛋白质，这些功能对于内含子的剪接和归巢/转座至关重要。这两个内含子都是高效的、位点特异性的逆转录元件，与非LTR逆转录转座子明显相关，包括人类基因组的LINE元件。主要和次要内含子归巢途径已经被鉴定，所有这些都依赖于一个显著的反应，其中含有内含子编码的RT和切除的内含子RNA酶的复合物通过反向剪接反应将内含子RNA插入无内含子DNA靶位点的有义链中。 后续步骤在各种RT非依赖性途径中差异很大。面包酵母具有许多属性，使其成为研究II组内含子归巢机制的强大而简单的系统，并且对于II组内含子的位点特异性归巢机制，为了清楚地了解各种途径如何不同，或者重要的是，为了显示共同归巢中间体如何在过程中分配，转座可能是内含子在进化过程中传播的机制。将开发一个系统来检测标记的内含子从线粒体到细胞核的逆转录转座。第三个目的是表征第二组内含子分子生物学的各个方面。主要的重点是识别核将分析基因，其处理内含子编码的RT的前体，控制切除的II组内含子RNA的稳定性和降解，并协助内含子的自我剪接。最后，将研究内含子编码蛋白的天然酶结构域的RNA结合和剪接功能。

项目成果

The stereochemical course of group II intron self-splicing.

II组内含子自剪接的立体化学过程。

• DOI: 10.1126/science.7527587
• 发表时间: 1994
• 期刊: Science (New York, N.Y.)
• 作者: Padgett,RA;Podar,M;Boulanger,SC;Perlman,PS
• 通讯作者: Perlman,PS

Mutant alleles of the MRS2 gene of yeast nuclear DNA suppress mutations in the catalytic core of a mitochondrial group II intron.

酵母核 DNA 的 MRS2 基因的突变等位基因抑制线粒体 II 组内含子催化核心的突变。

• DOI: 10.1006/jmbi.1998.2021
• 发表时间: 1998
• 期刊: Journal of molecular biology.
• 作者: Schmidt,U;Maue,I;Lehmann,K;Belcher,SM;Stahl,U;Perlman,PS
• 通讯作者: Perlman,PS

Reactions catalyzed by group II introns in vitro.

• DOI: 10.1016/s0076-6879(96)64010-5
• 发表时间: 1996-01-01
• 期刊: Methods in enzymology
• 作者: Perlman, P S;Podar, M
• 通讯作者: Podar, M

Mitochondrial gene expression in yeast: further studies of a self-splicing group II intron.

酵母中的线粒体基因表达：自剪接第二组内含子的进一步研究。

• DOI: 10.1007/978-1-4684-5251-8_4
• 发表时间: 1986
• 期刊: Basic life sciences
• 作者: Perlman,PS;Jarrell,KA;Dietrich,RC;Peebles,CL;Romiti,SL;Benatan,EJ
• 通讯作者: Benatan,EJ

Bacterial group II introns in a deep-sea hydrothermal vent environment.

深海热液喷口环境中的第二组细菌内含子。

• DOI: 10.1128/aem.68.12.6392-6398.2002
• 发表时间: 2002
• 期刊: Applied and environmental microbiology
• 影响因子: 4.4
• 作者: Podar,Mircea;Mullineaux,Lauren;Huang,Hon-Ren;Perlman,PhilipS;Sogin,MitchellL
• 通讯作者: Sogin,MitchellL