Development and mode of action of inhibitors of macromolecule biosynthesis from marine animals.

在我们从海洋生物中筛选海星（Asterina pectinifera）胚胎发育抑制剂的过程中，我们发现海洋海绵Rhopaloeides sp.的MeOH提取物在原肠胚形成之前的阶段特异性地抑制海星胚胎发生。初步检查导致分离出一种不寻常的降甾萜，命名为rhopaloic acid A，其抑制海星胚胎的原肠胚形成，并对人骨髓K-562细胞、人MOLT-4白血病细胞和小鼠L1210白血病细胞显示出强效细胞毒性7。对Rhopaloeides sp.提取物的进一步研究导致了相关代谢产物rhopaloic acids B和C的分离。我们确定了这些化合物的结构并揭示了生物活性。我们发现了一种新的核酸相关蛋白，命名为NAAP，它是从海星卵母细胞中分离的。用NAAP部分氨基酸序列的寡核苷酸克隆NAAP cDNA，并测定其全长cDNA序列。编码NAAP的转录本仅存在于生长的卵母细胞中，而不存在于完全生长的卵母细胞、胚胎和成体组织中。NAAP蛋白定位于成熟卵母细胞的萌发囊泡中，并在成熟卵母细胞减数分裂开始时萌发囊泡破裂后弥散分布。在卵母细胞成熟过程中，NAAP以细胞周期依赖的方式改变分子形式。结果表明，在芽囊泡期未被磷酸化的NAAP的Ser 145在减数分裂的M期被磷酸化。

During the course of our screening program for inhibitors of starfish (Asterina pectinifera) embryonic development from marine organisms, we found that the MeOH extract of the marine sponge Rhopaloeides sp. inhibited starfish embryogenesis specifically at the stage prior to gastrulation. A preliminary examination led to the isolation of an unusual norsesterterpene designated rhopaloic acid A which inhibited gastrulation of the starfish embryo and exhibited potent cytotoxicities against human myeloid K-562 cells, human MOLT-4 leukemia cells, and murine L1210 leukemia cells7. Further investigation of the extract of Rhopaloeides sp. led to the isolation of related metabolites, rhopaloic acids B and C.We determined the structure and uncovered biological activities of these compounds.We discovered a novel nucleic-acid associated protein designated as NAAP which was isolated from starfish oocytes. Oligonucleotides derived from the partial amino acid sequences of NAAP were used to clone NAAP cDNA and the entire cDNA sequence was determined. The transcript encoding NAAP was present only in growing oocytes but not in full-grown oocytes, embryos and adult tissues. The NAAP protein localized in the germinal vesicle of the full-grown oocyte and dispersed diffusely after germinal vesicle breakdown at the onset of meiosis. During oocyte maturation, NAAP changed the molecular form in a cell-cycle dependent manner. It was found that Ser 145 of NAAP, which was unphosphorylated at the germinal vesicle stage, was phosphorylated during the M phase of meiosis.