Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Japan.
Dev Genes Evol. 2011 Aug;221(3):157-66. doi: 10.1007/s00427-011-0368-2. Epub 2011 Jun 27.
Sulfatases such as arylsulfatase and heparan sulfate 6-O-endosulfatase play important roles in morphogenesis during sea urchin development. For the activation of these sulfatases, Cα-formylglycine formation by sulfatase modifying factor (Sumf) is required. In this study, to clarify the regulatory mechanisms for the activation of sulfatases during sea urchin development, we examined the expression and function of the Hemicentrotus pulcherrimus homologs of Sumf1 and Sumf2 (HpSumf1 and HpSumf2, respectively). Expression of HpSumf1 but not HpSumf2 mRNA was dynamically changed during early development. Functional analyses of recombinant HpSumf1 and HpSumf2 using HEK293T cells expressing mouse arylsulfatase A (ArsA) indicated that HpSumf1 and HpSumf2 were both able to activate mammalian ArsA. Knockdown of HpSumf1 using morpholino antisense oligonucleotides caused abnormal spicule formation in the sea urchin embryo. Injection of HpSumf2 mRNA had no effect on skeletogenesis, while injection of HpSumf1 mRNA induced severe supernumerary spicule formation. Taken together, these findings suggest that HpSumf1 is involved in the activation of sulfatases required for control of skeletogenesis.
硫酸酯酶，如芳基硫酸酯酶和肝素硫酸 6-O-内切硫酸酯酶，在海胆发育过程中的形态发生中发挥重要作用。对于这些硫酸酯酶的激活，需要硫酸酯酶修饰因子（Sumf）形成 Cα-甲酰甘氨酸。在这项研究中，为了阐明海胆发育过程中硫酸酯酶激活的调控机制，我们检查了 Hemicentrotus pulcherrimus 同源物 Sumf1 和 Sumf2（分别为 HpSumf1 和 HpSumf2）的表达和功能。在早期发育过程中，HpSumf1 的 mRNA 表达而不是 HpSumf2 的 mRNA 表达发生了动态变化。使用表达小鼠芳基硫酸酯酶 A（ArsA）的 HEK293T 细胞对重组 HpSumf1 和 HpSumf2 进行功能分析表明，HpSumf1 和 HpSumf2 都能够激活哺乳动物 ArsA。使用针对 HpSumf1 的 morpholino 反义寡核苷酸进行 HpSumf1 的敲低导致海胆胚胎中异常的骨针形成。HpSumf2 mRNA 的注射对骨骼发生没有影响，而 HpSumf1 mRNA 的注射诱导严重的额外骨针形成。总之，这些发现表明 HpSumf1 参与了硫酸酯酶的激活，这对于控制骨骼发生是必需的。
