Byrum C A, Walton K D, Robertson A J, Carbonneau S, Thomason R T, Coffman J A, McClay D R
Developmental, Cell, and Molecular Biology Group, Box 91000, Duke University, Durham, NC 27708, USA.
Dev Biol. 2006 Dec 1;300(1):194-218. doi: 10.1016/j.ydbio.2006.08.050. Epub 2006 Aug 25.
Protein phosphatases, in coordination with protein kinases, play crucial roles in regulation of signaling pathways. To identify protein tyrosine phosphatases (PTPs) and serine-threonine (ser-thr) phosphatases in the Strongylocentrotus purpuratus genome, 179 annotated sequences were studied (122 PTPs, 57 ser-thr phosphatases). Sequence analysis identified 91 phosphatases (33 conventional PTPs, 31 dual specificity phosphatases, 1 Class III Cysteine-based PTP, 1 Asp-based PTP, and 25 ser-thr phosphatases). Using catalytic sites, levels of conservation and constraint in amino acid sequence were examined. Nine of 25 receptor PTPs (RPTPs) corresponded to human, nematode, or fly homologues. Domain structure revealed that sea urchin-specific RPTPs including two, PTPRLec and PTPRscav, may act in immune defense. Embryonic transcription of each phosphatase was recorded from a high-density oligonucleotide tiling microarray experiment. Most RPTPs are expressed at very low levels, whereas nonreceptor PTPs (NRPTPs) are generally expressed at moderate levels. High expression was detected in MAP kinase phosphatases (MKPs) and numerous ser-thr phosphatases. For several expressed NRPTPs, MKPs, and ser-thr phosphatases, morpholino antisense-mediated knockdowns were performed and phenotypes obtained. Finally, to assess roles of annotated phosphatases in endomesoderm formation, a literature review of phosphatase functions in model organisms was superimposed on sea urchin developmental pathways to predict areas of functional activity.
蛋白磷酸酶与蛋白激酶协同作用,在信号通路调节中发挥关键作用。为了鉴定紫海胆基因组中的蛋白酪氨酸磷酸酶(PTP)和丝氨酸 - 苏氨酸(ser - thr)磷酸酶,研究了179个注释序列(122个PTP,57个ser - thr磷酸酶)。序列分析鉴定出91种磷酸酶(33种传统PTP、31种双特异性磷酸酶、1种III类基于半胱氨酸的PTP、1种基于天冬氨酸的PTP和25种ser - thr磷酸酶)。利用催化位点,检测了氨基酸序列中的保守程度和限制水平。25种受体PTP(RPTP)中有9种与人类、线虫或果蝇的同源物相对应。结构域结构显示,包括PTPRLec和PTPRscav在内的海胆特异性RPTP可能在免疫防御中发挥作用。通过高密度寡核苷酸平铺微阵列实验记录了每种磷酸酶的胚胎转录情况。大多数RPTP表达水平极低,而非受体PTP(NRPTP)通常以中等水平表达。在丝裂原活化蛋白激酶磷酸酶(MKP)和众多ser - thr磷酸酶中检测到高表达。对于几种表达的NRPTP、MKP和ser - thr磷酸酶,进行了吗啉代反义介导的敲低并获得了表型。最后,为了评估注释的磷酸酶在内胚层形成中的作用,将模型生物中磷酸酶功能的文献综述叠加到海胆发育途径上,以预测功能活性区域。