Screening and SNP mapping of copper-resistant mutations in C. elegans
Received date: 2014-05-12
Online published: 2014-12-20
Copper plays critical roles in biological system; however, it is toxic in excess. To identify novel genes involved in copper metabolism, we performed a whole genome-wide genetic screen in C. elegans model organism to search for mutants which are resistant to excessive copper. Wild type (N2) L4 worms were mutagenized with ethylmethane sulfonate (EMS), and the F2 progeny were screened on culture medium with excess copper. Two copper-resistant mutants, ms1 and ms2, were recovered from the screening of 100 000 hyploid genomes. No obvious developmental defects were observed in ms1 and ms2 mutants, and they were able to grow into adults on screen medium plate, but N2 worms arrested in L1 stage. Results of backcross test suggested that copper-resistant phenotype in ms1 may be controlled by a single recessive gene, but probably there are mutations in multiple genes in ms2, as no copper resistant worms could be found in F2 progeny when ms2 mutants were backcrossed with N2 worms. To determine the mutation positions of ms1, we employed single nucleotide polymorphisms (SNPs) mapping. Our mapping results indicated that ms1 mutation is on chromosome II (LGII). By analysis of 8 SNP markers from -18 to 23 on LGII, we found that ms1 mutation is at approximately LGII:-6. Further study on ms1 mutants will provide insights into copper metabolism and its regulation.
Shaojuan Song, Yaping Guo, Xueyao Zhang, Jianzhen Zhang, Enbo Ma . Screening and SNP mapping of copper-resistant mutations in C. elegans[J]. Hereditas(Beijing), 2014 , 36(12) : 1261 -1268 . DOI: 10.3724/SP.J.1005.2014.1261
[1] 朱志兀, 姚琳. 铜离子稳态平衡分子机理研究进展. 生命科学, 2012, 24(8): 847–857.
[2] 陈丽娟, 鲍远程, 余元勋, 关婷, 方长水. 铜转运分子机制及与Wilson病、Menkes病关系的研究进展. 现代中西医结合杂志, 2009, 18(32): 4036–4038.
[3] Gonzalez AG, Shirokova LS, Pokrovsky OS, Emnova EE, Martínez RE, Santana-Casiano JM, González-Dávila M, Pokrovski GS. Adsorption of copper on Pseudomonas au-reofaciens: protective role of surface exopolysaccharides. J Colloid Interface Sci, 2010, 350(1): 305–314.
[4] Bondarczuk K, Piotrowska-Seget Z. Molecular basis of active copper resistance mechanisms in Gram-negative bacteria. Cell Biol Toxicol, 2013, 29(6): 397–405.
[5] 丁艳菲, 王光钺, 傅亚萍, 朱诚. miR398在植物逆境胁迫应答中的作用. 遗传, 2010, 32(2): 129–134.
[6] 秦峰松, 杨崇林. 小线虫, 大发现: Caenorhabditis elegans在生命科学研究中的重要贡献. 生命科学, 2006, 18(5): 419–424.
[7] Wakabayashi T, Nakamura N, Sambongi Y, Wada Y, Oka T, Futai M. Identification of the copper chaperone, CUC-1, in Caenorhabditis elegans: tissue specific co-expression with the copper transporting ATPase, CUA-1. FEBS Lett, 1998, 440(1–2): 141–146.
[8] Calafato S, Swain S, Hughes S, Kille P, Stürzenbaum SR. Knock down of Caenorhabditis elegans cutc-1 exacerbates the sensitivity toward high levels of copper. Toxicol Sci, 2008, 106(2): 384–391. [9] Shaye DD, Greenwald I. OrthoList: A Compendium of C. elegans Genes with Human Orthologs. PLoS ONE, 2011, 6(5): e20085.
[10] Yoshimizu T, Omote H, Wakabayashi T, Sambongi Y, Futai M. Essential Cys-Pro-Cys motif of Caenorhabditis el-egans copper transport ATPase. Biosci Biotechnol Biochem, 1998, 62(6): 1258–1260.
[11] Sambongi Y, Wakabayashi T, Yoshimizu T, Omote H, Oka T, Futai M. Caenorhabditis elegans cDNA for a Menkes/ Wilson disease gene homologue and its function in a yeast CCC2 gene deletion mutant. J Biochem, 1997, 121(6): 1169–1175.
[12] Schwartz MS, Benci JL, Selote DS, Sharma AK, Chen AG, Dang H, Fares H, Vatamaniuk OK. Detoxification of mul-tiple heavy metals by a half-molecule ABC transporter, HMT-1, and coelomocytes of Caenorhabditis elegans. PLoS ONE, 2010, 5(3): e9564.
[13] Barsyte D, Lovejoy DA, Lithgow GJ. Longevity and heavy metal resistance in daf-2 and age-1 long-lived mutants of Caenorhabditis elegans. FASEB J, 2001, 15(3): 627–634.
[14] Song S, Guo Y, Zhang X, Zhang X, Zhang J, Ma E. Changes to cuticle surface ultrastructure and some biological func-tions in the nematode Caenorhabditis elegans exposed to excessive copper. Arch Environ Contam Toxicol, 2014, 66(3): 390–399.
[15] 赵淑清. 一种筛选拟南芥突变体的有效方法. 遗传, 2001, 23(3): 260–262.
[16] Brenner S. The genetics of Caenorhabditis elegans. Ge-netics, 1974, 77(1): 71–94.
[17] 贾熙华, 曹诚. 秀丽隐杆线虫在医药学领域的应用和进展. 药学学报, 2009, 44(7): 687–694.
[18] Jorgensen EM, Mango SE. The art and design of genetic screens: Caenorhabditis elegans. Nat Rev Genet, 2002, 3(5): 356–369.
[19] 秘彩莉, 沈银柱, 黄占景, 何聪芬, 柏峰, 马闻师, 赵宝存, 葛荣朝. 小麦耐盐突变体的分子生物学鉴定. 遗传, 1999, 21(6): 32–36.
[20] 刘喜冬, 王志鹏, 樊惠中, 李俊雅, 高会江. 基于高密度SNP标记的肉牛人工选择痕迹筛查. 遗传, 2012, 34(10): 1304–1313.
[21] 马云龙, 张勤, 丁向东. 利用高密度SNP检测不同猪品种间X染色体选择信号. 遗传, 2012, 34(10): 1251–1260.
[22] Davis MW, Hammarlund M, Harrach T, Hullett P, Olsen S, Jorgensen EM. Rapid single nucleotide polymorphism mapping in C. elegans. BMC Genomics, 2005, 6(1): 118.
[23] Swan KA, Curtis DE, McKusick KB, Voinov AV, Mapa FA, Cancilla MR. High-throughput gene mapping in Caeno-rhabditis elegans. Genome Res, 2002, 12(7): 1100–1105.
[24] 线虫基因数据库网站: www.wormbase.org.
[25] Hodgkin J. Introduction to genetics and genomics. In: WormBook. The C. elegans Research Community, Worm-Book. http://www.wormbook.org.
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