Wessel G M, Clark F, Berg L
Department of Molecular and Cellular Biology and Biochemistry, Brown University, Providence, RI 02912, USA.
Comp Biochem Physiol B Biochem Mol Biol. 1995 Mar;110(3):493-502. doi: 10.1016/0305-0491(94)00212-d.
Reversible tyrosine phosphorylation is involved in the fertilization reaction and early embryogenesis of the sea urchin (Foltz and Shilling, 1993; Ramachandran et al., 1993). To determine the enzymes present that may be involved in this regulation, we used a PCR screen to identify sequences that encode protein tyrosine kinases (PTK) and protein tyrosine phosphatases (PTP). We identified five PTKs and eight PTPs using cDNA libraries from two sea urchin species at two different stages of development, and the similarities to known PTK and PTP amino acid sequences ranged from 70 to 95%. The cognate proteins represented both "receptor"-class and cytoplasmic enzymes. Using RNAse protection assays we found that the respective mRNAs showed many accumulation profiles that we have grouped into three basic patterns: (1) mRNA levels that do not vary by more than two to three times throughout development; (2) mRNA levels highest in eggs or ovaries; and (3) mRNA levels highest in gastrula or pluteus stages. mRNAs specific to adult somatic cells of the ovary were not found, nor were mRNAs that accumulated selectively at the blastula stage. The results show that a diversity of enzymes involved in the regulation of reversible tyrosine phosphorylation is present in eggs and embryos of the sea urchin and that the differential accumulation in development of each mRNA suggests specific functional responsibilities by members of these enzyme families.
可逆性酪氨酸磷酸化参与海胆的受精反应和早期胚胎发育过程(福尔茨和希林,1993年;拉马钱德兰等人,1993年)。为了确定可能参与这种调节作用的酶,我们利用聚合酶链反应(PCR)筛选来鉴定编码蛋白酪氨酸激酶(PTK)和蛋白酪氨酸磷酸酶(PTP)的序列。我们使用来自两种海胆在两个不同发育阶段的cDNA文库,鉴定出了5种PTK和8种PTP,它们与已知PTK和PTP氨基酸序列的相似性在70%至95%之间。同源蛋白包括“受体”类和细胞质酶。通过核糖核酸酶保护试验,我们发现各自的mRNA呈现出许多积累模式,我们将其归为三种基本类型:(1)在整个发育过程中mRNA水平变化不超过两到三倍;(2)mRNA水平在卵或卵巢中最高;(3)mRNA水平在原肠胚或长腕幼虫阶段最高。未发现卵巢成年体细胞特有的mRNA,也未发现只在囊胚期选择性积累的mRNA。结果表明,海胆的卵和胚胎中存在多种参与可逆性酪氨酸磷酸化调节的酶,并且每种mRNA在发育过程中的差异积累表明这些酶家族成员具有特定的功能作用。