Smolenicka Zuzana, Pani Filippo, Hwang Kae-Jung, Gruschus James M, Ferretti James A
International Marine Centre, Localita Sa Mardini Torregrande (OR), Sardinia 09072, Italy.
Cell Biol Int. 2003;27(2):81-7. doi: 10.1016/s1065-6995(02)00351-7.
The partial sequence of a novel homeobox-containing gene from Paracentrotus lividus is described. Both cDNA and genomic DNA were screened using probes from the vnd/NK-2 homeobox gene found in Drosophila melanogaster. The new DNA sequence found in P. lividus encodes a protein fragment that is closely related to the NK family of homeodomain transcriptional regulators originally discovered in the fruit fly. This study thus represents the first finding of a homeobox gene from the NK family in sea urchin. The DNA that was sequenced includes the most highly conserved region of the NK genes and contains the 180 basepair homeobox (i.e. the DNA segment that encodes the homeodomain), the NK-2 box that encodes the NK-2-specific domain (NK-2 SD), and the acidic box that encodes an acidic domain, but which is found only in a limited subset of the NK genes. In this deduced sequence, the 60 amino acid residue homeodomain contains tyrosine in position 54 and leucine in position 7, which implies that the protein will bind to an unusual sequence of DNA that contains 5'-CAAGTG-3' as its core. The presence of tyrosine in position 54 identifies the gene as a member of the NK-2 class of homeobox genes. Positions 37 and 56 of the homeodomain contain isoleucine and leucine, respectively, which is the first finding in the NK family of homeodomains of these particular amino acid residues in those positions. The presence of the NK-2 box is consistent with identification of the gene as a member of the NK-2 class, and suggests an important role for the C-terminal portion of the protein in transcriptional activation. The sequence homology of the NK-2 box and the spacing between it and the homeobox further suggest that this gene is a member of the NKx-2.2 subclass, whose genes typically are expressed in brain and play a role in axonal guidance, and whose full lengths often are of the order of 900 bases. Homologous NK genes have been found in such diverse invertebrate and vertebrate species, such as Amphioxus sp., Xenopus sp., Caenorhabditis elegans, zebra fish, chicken, hamster, mouse and humans. The finding of this new gene together with sequence comparisons suggests possible evolutionary relationships between sea urchins and vertebrates in the developmental pathways of their body plans.
本文描述了来自紫球海胆(Paracentrotus lividus)的一个新的含同源异型框基因的部分序列。利用从黑腹果蝇(Drosophila melanogaster)中发现的vnd/NK-2同源异型框基因的探针筛选了互补DNA(cDNA)和基因组DNA。在紫球海胆中发现的新DNA序列编码一个与最初在果蝇中发现的同源异型域转录调节因子NK家族密切相关的蛋白质片段。因此,本研究代表了在海胆中首次发现NK家族的同源异型框基因。测序的DNA包括NK基因中最保守的区域,包含180个碱基对的同源异型框(即编码同源异型域的DNA片段)、编码NK-2特异性结构域(NK-2 SD)的NK-2框以及编码酸性结构域的酸性框,但酸性框仅在NK基因的有限子集中发现。在这个推导序列中,60个氨基酸残基的同源异型域在第54位含有酪氨酸,在第7位含有亮氨酸,这意味着该蛋白质将与以5'-CAAGTG-3'为核心的异常DNA序列结合。第54位存在酪氨酸将该基因鉴定为同源异型框基因NK-2类的成员。同源异型域的第37位和第56位分别含有异亮氨酸和亮氨酸,这是在NK家族同源异型域中首次在这些位置发现这些特定氨基酸残基。NK-2框的存在与将该基因鉴定为NK-2类成员一致,并表明该蛋白质的C末端部分在转录激活中起重要作用。NK-2框的序列同源性及其与同源异型框之间的间距进一步表明该基因是NKx-2.2亚类的成员,其基因通常在大脑中表达并在轴突导向中起作用,其全长通常约为900个碱基。在多种无脊椎动物和脊椎动物物种中都发现了同源的NK基因,如文昌鱼(Amphioxus sp.)、非洲爪蟾(Xenopus sp.)、秀丽隐杆线虫(Caenorhabditis elegans)、斑马鱼、鸡、仓鼠、小鼠和人类。这个新基因的发现以及序列比较表明,海胆和脊椎动物在其身体结构的发育途径中可能存在进化关系。