SUMO proteomics in C. elegans; germ line, meiosis and the DNA damage response

线虫中的相扑蛋白质组学；

• 批准号: BB/J015199/1
• 负责人: Anton Gartner
• 金额: $ 38.62万
• 依托单位: University of Dundee
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FJ015199%2F1
• 关键词: SUMO; proteomics; C.; elegans; germ

Proteins need to be regulated to properly fulfil their function in cells and living organisms. Specialized sets of proteins are required to convey a variety of signals within living cells. Such signalling proteins confer signals by attaching distinct sets of molecules to their target proteins. The attachment of such molecules, referred to as modification serves as a 'cellular language'. Modern molecular biology research focuses on such signalling proteins and tries to address their importance in modifying the activity of their targets. Modifications that confer signals such as phosphorylation or ubiquitylation have been known for a long time. However, only in the past decade, it became clear that sumoylation provides an important way of intracellular communication. The Hay laboratory elaborated important molecular details pertaining to basic biochemical reactions that are needed for SUMO modification. SUMO by itself is a sizable protein, that is attached to certain residues of its target proteins and several proteins are needed to confer sumoylation. So called 'systems approaches' aim to provide a comprehensive analysis and understanding of the 'SUMO language'. Such analysis requires the identification, of all SUMO modified proteins, and a detailed analysis of how and to which extend SUMO modification changes in response to various physiological and pathophysiological conditions. First experiments to assess the global role of SUMO modification were done in with cell cultures. While those systems have obvious experimental advantages, these cells do not necessarily reflect the behaviour of cells in intact organisms. This is why we plan to use the small roundworm C. elegans as a model system to globally asses the extend, and importance of sumoylation in a living organism. C. elegans was chosen as the simplest possible animal system. Studies are facilitated by the simplicity of the organism at the developmental and anatomical level, by the ease of its maintenance, as well as by the availability of methods that allow for manipulating its genetic information procedures necessary to study the importance of SUMO modification. Despite of its simplicity, C. elegans is a multicellular organism that shares many fundamental genetic programs with humans. Thus results obtained in the C. elegans system are likely to be applicable to humans and working with worms does not raise ethical concerns. As part of our studies we want to express modified versions of SUMO in the worm, such that we can best apply the advanced methods we have developed to isolate all SUMO modified proteins. To identify those proteins, but also to assess the extent of SUMO modification we will use a method we recently developed allowing for the identification and quantification of thousands of worm proteins. We not only want to find all SUMO modified proteins, but also want to assess the importance of SUMO modification. To start with this, we chose an important biological processes extensively studied in our labs. 1) Meiosis is a specialised cell division, needed for the generation of germ cells. During this division the duplicated set of chromosomes occurring in normal cells is reduced to just one copy. The subsequent fusion of germ cell restores a duplicated set of chromosomes one coming from the father, another one coming from the mother. Not only this, during meiosis a process referred to as recombination also mixes individual chromosomes such that chromosomes contain elements from both grandparents. Recombination ensures genetic diversity. 2) Recombination pertains to the breaking and linking DNA molecules. Recombination is used when DNA damaging agents such as ionizing irradiation breaks chromosomes. We already have know that SUMO modification is important for meiosis and recombination and we now want perform a systematic analysis of this. Our studies are important to understand how the genetic information is passed on from one generation to the next.

需要调节蛋白质以正确履行其在细胞和生物体中的功能。需要专门的蛋白质集来传达活细胞中的各种信号。这种信号蛋白通过将不同的分子集与其靶蛋白相结合来赋予信号。这种分子的附着，称为修饰，用作“蜂窝语言”。现代分子生物学研究的重点是这种信号蛋白，并试图解决它们在修改目标活性方面的重要性。赋予信号（例如磷酸化或泛素化）的修改已有很长时间了。但是，只有在过去的十年中，很明显，Sumoylation提供了一种重要的细胞内交流方式。干草实验室详细阐述了与Sumo修饰所需的基本生化反应有关的重要分子细节。 Sumo本身是一种相当大的蛋白质，它附着在其靶蛋白的某些残基上，需要几种蛋白来赋予Sumoylation。所谓的“系统方法”旨在提供对“ Sumo语言”的全面分析和理解。这样的分析需要对所有SUMO修饰蛋白的识别，以及详细的分析，分析如何以及向Sumo修饰的变化而变化，以响应各种生理和病理生理条件。在细胞培养物中进行了第一个评估Sumo修饰的全球作用的实验。尽管这些系统具有明显的实验优势，但这些细胞并不一定反映细胞在完整生物中的行为。这就是为什么我们计划将小round虫秀丽隐杆线虫作为模型系统，以在全球范围内扩展和在活生物体中的sumoylation的重要性。秀丽隐杆线虫被选为最简单的动物系统。由于生物体在发育和解剖学水平上的简单性，通过其维护的简化以及允许操纵其基因信息修改重要性所必需的遗传信息程序的可用性来促进研究。尽管它很简单，但秀丽隐杆线虫还是一种多细胞生物，与人类共享许多基本遗传程序。因此，在秀丽隐杆线虫系统中获得的结果可能适用于人类，并且与蠕虫一起工作不会引起道德问题。作为研究的一部分，我们希望在蠕虫中表达Sumo的修改版本，以便我们可以最好地应用我们开发的先进方法来隔离所有Sumo修饰的蛋白质。为了识别这些蛋白质，但为了评估Sumo修饰的程度，我们将使用最近开发的方法，允许识别和定量数千种蠕虫蛋白。我们不仅想找到所有Sumo修饰的蛋白质，而且还希望评估Sumo修饰的重要性。首先，我们选择了在实验室中广泛研究的重要生物学过程。 1）减数分裂是生成生殖细胞所需的专门细胞分裂。在此划分期间，在正常细胞中发生的重复的染色体将减少为仅一个拷贝。随后的生殖细胞融合恢复了一套重复的染色体，其中一个来自父亲，另一个来自母亲。不仅如此，在减数分裂过程中，称为重组的过程还将单个染色体混合在一起，因此染色体含有两个祖父母的元素。重组确保遗传多样性。 2）重组与断裂和连接DNA分子有关。当DNA损坏剂（例如电离辐射断裂）染色体时，使用重组。我们已经知道，Sumo修饰对于减数分裂和重组很重要，现在我们希望对此进行系统分析。我们的研究对于了解如何从一代传递到另一代的遗传信息很重要。

项目成果

The SMC-5/6 Complex and the HIM-6 (BLM) Helicase Synergistically Promote Meiotic Recombination Intermediate Processing and Chromosome Maturation during Caenorhabditis elegans Meiosis.

• DOI: 10.1371/journal.pgen.1005872
• 发表时间: 2016-03
• 期刊: PLoS genetics
• 影响因子: 4.5
• 作者: Hong Y;Sonneville R;Agostinho A;Meier B;Wang B;Blow JJ;Gartner A
• 通讯作者: Gartner A

LEM-3 is a midbody-tethered DNA nuclease that resolves chromatin bridges during late mitosis.

• DOI: 10.1038/s41467-018-03135-w
• 发表时间: 2018-02-20
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Hong Y;Sonneville R;Wang B;Scheidt V;Meier B;Woglar A;Demetriou S;Labib K;Jantsch V;Gartner A
• 通讯作者: Gartner A

Erratum: Dynamic SUMO modification regulates mitotic chromosome assembly and cell cycle progression in Caenorhabditis elegans.

• DOI: 10.1038/ncomms7352
• 发表时间: 2015-02-12
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Pelisch, Federico;Sonneville, Remi;Pourkarimi, Ehsan;Agostinho, Ana;Blow, J. Julian;Gartner, Anton;Hay, Ronald T.
• 通讯作者: Hay, Ronald T.

A SUMO-Dependent Protein Network Regulates Chromosome Congression during Oocyte Meiosis.

• DOI: 10.1016/j.molcel.2016.11.001
• 发表时间: 2017-01-05
• 期刊: Molecular cell
• 影响因子: 16
• 作者: Pelisch F;Tammsalu T;Wang B;Jaffray EG;Gartner A;Hay RT
• 通讯作者: Hay RT

Dynamic SUMO modification regulates mitotic chromosome assembly and cell cycle progression in Caenorhabditis elegans.

• DOI: 10.1038/ncomms6485
• 发表时间: 2014-12-05
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Pelisch, Federico;Sonneville, Remi;Pourkarimi, Ehsan;Agostinho, Ana;Blow, J. Julian;Gartner, Anton;Hay, Ronald T.
• 通讯作者: Hay, Ronald T.