Botany Department, La Trobe University, Melbourne, Victoria 3086, Australia.
Plant Cell. 2011 Jun;23(6):2209-24. doi: 10.1105/tpc.110.082651. Epub 2011 Jun 14.
Arabidopsis thaliana MYB80 (formerly MYB103) is expressed in the tapetum and microspores between anther developmental stages 6 and 10. MYB80 encodes a MYB transcription factor that is essential for tapetal and pollen development. Using microarray analysis of anther mRNA, we identified 404 genes differentially expressed in the myb80 mutant. Employing the glucocorticoid receptor system, the expression of 79 genes was changed when MYB80 function was restored in the myb80 mutant following induction by dexamethasone. Thirty-two genes were analyzed using chromatin immunoprecipitation, and three were identified as direct targets of MYB80. The genes encode a glyoxal oxidase (GLOX1), a pectin methylesterase (VANGUARD1), and an A1 aspartic protease (UNDEAD). All three genes are expressed in the tapetum and microspores. Electrophoretic mobility shift assays confirmed that MYB80 binds to all three target promoters, with the preferential binding site containing the CCAACC motif. TUNEL assays showed that when UNDEAD expression was silenced using small interfering RNA, premature tapetal and pollen programmed cell death occurred, resembling the myb80 mutant phenotype. UNDEAD possesses a mitochondrial targeting signal and may hydrolyze an apoptosis-inducing protein(s) in mitochondria. The timing of tapetal programmed cell death is critical for pollen development, and the MYB80/UNDEAD system may regulate that timing.
拟南芥 MYB80(原 MYB103)在花药发育阶段 6 到 10 期的绒毡层和小孢子中表达。MYB80 编码一个 MYB 转录因子,对绒毡层和花粉发育是必需的。通过花药 mRNA 的微阵列分析,我们鉴定了在 myb80 突变体中差异表达的 404 个基因。利用糖皮质激素受体系统,当 MYB80 功能在 dexamethasone 诱导下在 myb80 突变体中恢复时,79 个基因的表达发生改变。使用染色质免疫沉淀分析了 32 个基因,其中 3 个被鉴定为 MYB80 的直接靶标。这些基因编码一个乙醛氧化酶(GLOX1)、一个果胶甲酯酶(VANGUARD1)和一个 A1 天冬氨酸蛋白酶(UNDEAD)。所有这三个基因都在绒毡层和小孢子中表达。电泳迁移率变动分析证实 MYB80 与所有三个靶标启动子结合,优先结合位点含有 CCAACC 基序。TUNEL 分析表明,当使用小干扰 RNA 沉默 UNDEAD 表达时,过早的绒毡层和花粉程序性细胞死亡发生,类似于 myb80 突变体表型。UNDEAD 具有一个线粒体靶向信号,可能水解线粒体中的一个凋亡诱导蛋白。绒毡层程序性细胞死亡的时间对于花粉发育至关重要,而 MYB80/UNDEAD 系统可能调节该时间。