Stowers R S, Garza D, Rascle A, Hogness D S
Department of Biochemistry, Stanford University School of Medicine, Beckman Center, B300, Stanford, California 94305-5329, USA.
Dev Biol. 2000 May 1;221(1):23-40. doi: 10.1006/dbio.2000.9685.
The pulse of ecdysone that triggers Drosophila metamorphosis activates six early genes in a primary response made visible by polytene chromosome puffs. The secondary response is detected by the induction of over 100 late puffs, only a few of which have been subject to molecular genetic analysis. We present a molecular and mutational analysis of the L63 gene responsible for the late puff at 63E. This gene contains overlapping L63A, B, and C transcription units of which the A unit encodes two isoforms and the B unit three. The C unit, which exhibits little activity, encodes one of the B isoforms. Evidence that L63B, but not L63A, transcription is ecdysone responsive derives from their developmental transcription profiles and from P-element mutagenesis showing that ecdysone induction of the 63E puff requires sequences adjacent to the 5' end of L63B but not those adjacent to the 5' end of L63A. L63-specific lethal mutations showed that L63 is required not only for metamorphosis, but also maternally and for embryonic and larval development. The L63 proteins contain a common C-terminal 294-aa sequence that is 71% identical to the CDK sequence of the murine PFTAIRE protein. In vivo tests of L63 proteins altered by site-directed mutagenesis showed that they exhibit CDK functions. L63 proteins are widely distributed among late larval and prepupal tissues and are unlikely to be involved in cell cycle functions.
触发果蝇变态的蜕皮激素脉冲在多线染色体胀泡所显示的初级反应中激活六个早期基因。次级反应通过诱导100多个晚期胀泡来检测,其中只有少数几个已进行分子遗传学分析。我们对负责63E处晚期胀泡的L63基因进行了分子和突变分析。该基因包含重叠的L63A、B和C转录单位,其中A单位编码两种异构体,B单位编码三种。活性较低的C单位编码B异构体之一。L63B而非L63A的转录对蜕皮激素有反应的证据来自它们的发育转录谱以及P因子诱变,表明63E胀泡的蜕皮激素诱导需要L63B 5'端附近的序列,而不是L63A 5'端附近的序列。L63特异性致死突变表明,L63不仅对变态是必需的,而且对母体以及胚胎和幼虫发育也是必需的。L63蛋白含有一个共同的C末端294个氨基酸的序列,该序列与小鼠PFTAIRE蛋白的CDK序列有71%的同一性。对通过定点诱变改变的L63蛋白进行的体内测试表明它们具有CDK功能。L63蛋白广泛分布于晚期幼虫和蛹前期组织中,不太可能参与细胞周期功能。