Role of Signal-transduction in the Development of C.elegans

信号转导在秀丽隐杆线虫发育中的作用

• 批准号: 04454615
• 负责人: KATSURA Isao
• 金额: $ 3.97万
• 依托单位: National Institute of Genetics
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454615/
• 关键词: C.elegans; Signal transduction; Protein kinase; Fluoride ion; Mutants; クローニング; 耐性変異

The fluoride-resistant (flr) mutations of the nematode C.elegans are all recessive and consist of two classes. Class 1 mutations (flr-1, flr-3 and flr-4) confer strong resistance and slow growth on worms, whereas class 2 mutations (flr-2 and flr-5) confer weak resistance, keep the growth rate unchanged and suppress the slow-growing phenotype of class 1 mutations. In this study we mapped the class 2 mutations and made heterozygotes between either of these mutations and a deficiency including it. The phenotypes of the heterozygotes showed that all the class 2 mutations are hypomorphs or null mutations. We also performed complementation tests between the flr mutations and some known mutations, which confirmed that they map in new genes. The flr genes probably act in neural functions, because mutations in these genes affect formation of dauer larvae under certain circumstances. We cloned two of the flr genes, flr-1 and flr-3, and their cDNA (incomplete length). Sequencing studies showed th … More at the deduced amino acid sequence of flr-1 has weak homology to some ion channels, whereas flr-3 most probably codes for a protein kinase of a new family. Since we obtained two kinds of flr-3 cDNAs differing in the 5' region, the expression of flr-3 may be regulated by alternative splicing.The "clr-1-like" mutations are defined as larval lethal mutations that cause detachment of the outer surface of the intestine from the inner surface of the body wall. They map in known signal-transduction genes (let-23, let-341, let-60, lin-45, sem-5, lag-2, clr-1 etc.) and unknown genes. We think the latter genes are members of the signal-transduction systems, or they act in cell-differentiation or cell-functions downstream to the signal-transduction systems. In this study we cloned a 4.4kb DNA fragment that contains let(ut102), one of the unknown genes for the clr-1-like mutations. We are sequencing the clone, but homology to known genes has not been detected so far. We plan to study the position of expression of the gene in worms to look for the target of the lethal mutation. We will also search for the relation between the let(ut102) gene and known signal-transduction systems by checking change of its expression by signal-transduction mutations and by testing suppression of the mutations by microinjection of the clone into the mutant worms. Less

线虫的耐氟化氢(Flr)突变均为隐性突变，分为两类。1类突变(flr-1、flr-3和flr-4)赋予蠕虫较强的抗性和缓慢的生长，而2类突变(flr-2和flr-5)赋予蠕虫较弱的抵抗力，保持生长速度不变，并抑制1类突变的缓慢生长表型。在这项研究中，我们绘制了2类突变的图谱，并使这些突变中的任何一个与包括它的缺陷之间的杂合子。杂合子的表型分析表明，所有2类突变均为亚型突变或零突变。我们还进行了flr突变和一些已知突变之间的互补测试，证实它们映射到新的基因中。Flr基因可能在神经功能中起作用，因为这些基因的突变在某些情况下会影响达尔幼虫的形成。我们克隆了两个flr基因，flr-1和flr-3，以及它们的全长。测序研究表明…更重要的是，flr-1的推导氨基酸序列与一些离子通道有微弱的同源性，而flr-3很可能编码一个新的家族的蛋白激酶。由于我们获得了两种在5‘端不同的flr-3基因，flr-3的表达可能受到选择性剪接的调控。“类CLR-1”突变被定义为导致幼虫肠道外表面与体壁内表面分离的致命性突变。它们定位于已知的信号转导基因(let-23、let-341、let-60、lin-45、sem-5、Lag-2、clr-1等)。和未知的基因。我们认为后者是信号转导系统的成员，或者它们在信号转导系统下游的细胞分化或细胞功能中起作用。在这项研究中，我们克隆了一个4.4kb的DNA片段，其中包含LET(Ut102)，这是CLR-1样突变的未知基因之一。我们正在对克隆进行测序，但到目前为止还没有检测到与已知基因的同源性。我们计划研究该基因在蠕虫中的表达位置，以寻找致命突变的靶点。我们还将通过检查let(Ut102)基因在信号转导突变中表达的变化，以及通过将克隆显微注射到突变蠕虫体内来测试对突变的抑制，来寻找let(Ut102)基因与已知信号转导系统之间的关系。较少

项目成果

I.Katsura et al.: "More Mutants from Mishima" The Worm Breeder's Gazette. 12-4. 66 (1992)

I.Katsura 等人：“来自三岛的更多突变体”蠕虫饲养者公报。

I.Katsura: "Genes acting in Morphogenesis of the Nemtode C.elegans (in Japanese)" Jikkenn Igaku (Experimental Medicine). 11-12. 1570-1576 (1994)

I.Katsura：“影响线虫形态发生的基因（日语）”Jikkenn Igaku（实验医学）。

I.Katsura: "In Search of New Mutants in Cell-signaling Systems of the Nematode Caenorhabditis elegans." Genetica. 88. 137-146 (1993)

I.Katsura：“寻找线虫细胞信号系统中的新突变体。”

I.Katura: "In search of new mutants in cell-signdlling systems of the nematode Caenorhabditis elegans." Genetica. (1993)

I.Katura：“在线虫细胞信号系统中寻找新的突变体。”

I.Katsura et al.: "Isolation,Characterization,and Epistasis of Fluoride‐resistant Mrtants of Caenorhabdetis elegans." Genetics. 136. 145-154 (1994)

I. Katsura 等人：“秀丽隐杆线虫耐氟化物的分离、表征和上位性”，136. 145-154 (1994)。