Essential roles of 3′-phosphoadenosine 5′-phoshosulfate synthase in embryonic and larval development of the nematode Caenorhabditis elegans
Essential roles of 3′-phosphoadenosine 5′-phoshosulfate synthase in embryonic and larval development of the nematode Caenorhabditis elegans
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DOI：
10.1074/jbc.m601509200
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发表时间：
2006-04-21
影响因子：
4.8
通讯作者：
Nomura, K
中科院分区：
文献类型：
--
作者：
Dejima, K;Seko, A;Nomura, K
生物分子的硫酸化在许多生物过程中起着关键作用，从细菌到人类都广泛存在。所有生物体中的所有硫酸化反应都需要活化的硫酸盐，3 '-磷酸腺苷5'-磷酸硫酸盐（PAPS）作为通用供体。在动物中，PAPS由ATP和无机硫酸盐通过双功能酶PAPS合成酶合成。在哺乳动物中，PAPS合成酶2（两种PAPS合成酶同工酶之一）的遗传缺陷导致侏儒症，但对PAPS合成完全丧失的后果知之甚少。为了确定硫酸化的发展作用，我们克隆了秀丽隐杆线虫PAPS酶同源基因，pps-1，并通过分离缺失突变体和RNA介导的干扰耗尽其产物的表达。PPS-1蛋白在体外表现出形成PAPS的特异性活性，并且通过RNAi破坏pps-1基因导致肌肉图案化和上皮细胞形状变化中的多效性发育缺陷，伴随糖胺聚糖硫酸化的减少。此外，pps-1无效突变体表现出幼虫致死性。这些数据表明，硫酸化是必不可少的正常生长和表皮的完整性，在C。优美的此外，报告基因分析显示，pps-1在表皮和几种腺细胞中表达，但在神经元和肌肉中不表达，表明神经元和肌肉中的PAPS由其他细胞提供。
Sulfation of biomolecules, which is widely observed from bacteria to humans, plays critical roles in many biological processes. All sulfation reactions in all organisms require activated sulfate, 3'-phosphoadenosine 5'-phosphosulfate ( PAPS), as a universal donor. In animals, PAPS is synthesized from ATP and inorganic sulfate by the bifunctional enzyme, PAPS synthase. In mammals, genetic defects in PAPS synthase 2, one of two PAPS synthase isozymes, cause dwarfism disorder, but little is known about the consequences of the complete loss of PAPS synthesis. To define the developmental role of sulfation, we cloned a Caenorhabditis elegans PAPS synthase-homologous gene, pps-1, and depleted expression of its product by isolating the deletion mutant and by RNA-mediated interference. PPS-1 protein exhibits specific activity to form PAPS in vitro, and disruption of the pps-1 gene by RNAi causes pleiotropic developmental defects in muscle patterning and epithelial cell shape changes with a decrease in glycosaminoglycan sulfation. Additionally, the pps-1 null mutant exhibits larval lethality. These data suggest that sulfation is essential for normal growth and integrity of epidermis in C. elegans. Furthermore, reporter analysis showed that pps-1 is expressed in the epidermis and several gland cells but not in neurons and muscles, indicating that PAPS in the neurons and muscles is provided by other cells.