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Positive regulation of minichromosome maintenance gene expression DNA replication and cell transformation by a plant retinoblastoma gene

机译:植物视网膜母细胞瘤基因对微染色体维持基因表达DNA复制和细胞转化的正调控

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摘要

Retinoblastoma-related (RBR) genes inhibit the cell cycle primarily by repressing adenovirus E2 promoter binding factor (E2F) transcription factors, which drive the expression of numerous genes required for DNA synthesis and cell cycle progression. The RBR-E2F pathway is conserved in plants, but cereals such as maize are characterized by having a complex RBR gene family with at least 2 functionally distinct members, RBR1 and RBR3. Although RBR1 has a clear cell cycle inhibitory function, it is not known whether RBR3 has a positive or negative role. By uncoupling RBR3 from the negative regulation of RBR1 in cultured maize embryos through a combination of approaches, we demonstrate that RBR3 has a positive and critical role in the expression of E2F targets required for the initiation of DNA synthesis, DNA replication, and the efficiency with which transformed plants can be obtained. Titration of endogenous RBR3 activity through expression of a dominant-negative allele with a compromised pocket domain suggests that these RBR3 functions require an activity distinct from its pocket domain. Our results indicate a cell cycle pathway in maize, in which 2 RBR genes have specific and opposing functions. Thus, the paradigm that RBR genes are negative cell cycle regulators cannot be considered universal.
机译:视网膜母细胞瘤相关(RBR)基因主要通过抑制腺病毒E2启动子结合因子(E2F)转录因子来抑制细胞周期,该因子驱动DNA合成和细胞周期进程所需的许多基因的表达。 RBR-E2F途径在植物中是保守的,但是谷物(例如玉米)的特征是具有复杂的RBR基因家族,该家族具有至少2个功能不同的成员RBR1和RBR3。尽管RBR1具有明确的细胞周期抑制功能,但不知道RBR3是发挥积极作用还是消极作用。通过多种方法的结合,将RBR3与培养的玉米胚中RBR1的负调控脱钩,我们证明RBR3在E2F靶标的表达中具有积极和关键的作用,而E2F靶标对于DNA合成,DNA复制的启动以及可以获得哪些转化植物。通过表达具有受损口袋结构域的显性阴性等位基因来内源性RBR3活性的滴定表明,这些RBR3功能需要与其口袋结构域不同的活性。我们的结果表明了玉米中的细胞周期途径,其中2个RBR基因具有特异性和相反的功能。因此,RBR基因是负细胞周期调节因子的范式不能被认为是普遍的。

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