Sano T, Tabuse Y, Nishiwaki K, Miwa J
Fundamental Research Laboratories, NEC Corporation, Tsukuba, Japan.
J Mol Biol. 1995 Aug 25;251(4):477-85. doi: 10.1006/jmbi.1995.0449.
The gene tpa-1 on chromosome IV of the nematode Caenorhabditis elegans plays a major and definitive role in the adversary action of tumour-promoting phorbol esters, which induce growth arrest and locomotory distress in the animal. The gene was deduced to code for a protein kinase C (PKC) homologue by molecular cloning. We have now sequenced the complete genomic and complementary DNAs for tpa-1 and have analysed their structural features in detail: (1) tpa-1 spans over 20 kb consisting of eleven exons and ten introns; (2) two different-sized mRNAs are generated from the tpa-1 locus; (3) both mRNAs are trans-spliced to the trans-spliced leader SL1; (4) both mRNAs encode PKC isoforms, which are most similar to Ca(2+)-independent novel PKC0; (5) the two PKC isoforms differ from each other in that the smaller lacks the amino-terminal region of the larger corresponding to the first four exons and a portion of the fifth exon; and (6) three introns are located at; identical positions in the polypeptide sequences aligned between the C. elegans tpa-1 product and a PKC of the fruit fly Drosophila melanogaster.
线虫秀丽隐杆线虫IV号染色体上的基因tpa-1在促肿瘤佛波酯的对抗作用中发挥着主要且决定性的作用,促肿瘤佛波酯会导致该动物生长停滞和运动障碍。通过分子克隆推断该基因编码一种蛋白激酶C(PKC)同源物。我们现已对tpa-1的完整基因组DNA和互补DNA进行了测序,并详细分析了它们的结构特征:(1)tpa-1跨度超过20 kb,由11个外显子和10个内含子组成;(2)tpa-1基因座产生两种不同大小的mRNA;(3)两种mRNA都反式剪接到反式剪接前导序列SL1;(4)两种mRNA都编码PKC同工型,它们与不依赖Ca(2+)的新型PKC0最为相似;(5)两种PKC同工型的不同之处在于,较小的同工型缺少较大同工型对应于前四个外显子和第五个外显子一部分的氨基末端区域;(6)三个内含子位于秀丽隐杆线虫tpa-1产物与果蝇黑腹果蝇的PKC之间比对的多肽序列中的相同位置。