Lebedeva L I, Dubatolova T D, Omel'ianchuk L V
Tsitologiia. 2013;55(6):406-13.
Mitosis, cytokinesis and nuclear texture of wing imaginal discs cells silenced by UAS-RNAi-dlg construct induced by 1096-Ga 14 driver were studied. The silencing construct contains coding region of dlg gene and the complementary region. Further, this RNA hairpin (Dietzl et al., 2007) is processed by endogenous protein Dicer and the resulting RNA fragments silence mRNA dlg. Tumor suppressor gene dlg is encoding for 21 transcripts. The construct UAS-RNAi-dlg inactivates 14 transcripts--RE, RH, RQ, RS, RG, RD, RL, RB, RK, RR, RT, RN, RA, RP--and does not silenced the other 7 (RO, RF, RI, RU, RJ, RC, RM). This permits to study functions of proteins containg guanilate-kinase domain IPR008145 at C-end of the protein. The most important consequences of the silencing are abnormal mitotic exit and the formation of binuclear cells. Quantitative fluorescence measurements of anti-H3-p histone and DAPI signals showed phase-specific changes in nuclear texture. The inactivation of cellular cortex polarization is the most likely target of dlg inactivation in mitosis.
研究了由1096 - Ga 14驱动子诱导的UAS - RNAi - dlg构建体沉默的翅成虫盘细胞的有丝分裂、胞质分裂和核纹理。该沉默构建体包含dlg基因的编码区和互补区。此外,这种RNA发夹(Dietzl等人,2007)由内源性蛋白质Dicer加工,产生的RNA片段使dlg mRNA沉默。肿瘤抑制基因dlg编码21种转录本。UAS - RNAi - dlg构建体使14种转录本失活——RE、RH、RQ、RS、RG、RD、RL、RB、RK、RR、RT、RN、RA、RP——而不会使其他7种(RO、RF、RI、RU、RJ、RC、RM)沉默。这使得能够研究在蛋白质C端含有鸟苷酸激酶结构域IPR008145的蛋白质的功能。沉默的最重要后果是有丝分裂异常退出和双核细胞的形成。抗H3 - p组蛋白和DAPI信号的定量荧光测量显示核纹理存在阶段特异性变化。细胞皮质极化的失活最有可能是有丝分裂中dlg失活的靶点。