Biochemistry and Function of C.elegans RNA Helicase A

描述（由申请人提供）：生物体以精确组织的方式发育 由复杂的细胞机制控制的模式。许多这些 在高等真核生物中是保守的。一些蛋白质 负责指导身体特定部位的发育（例如， 同源异型蛋白）。其他如RNA解旋酶A（RHA），则参与基本的 所有细胞中的细胞过程，这些蛋白质的缺陷产生 整个生物体的发育异常。RNA解旋酶参与 在mRNA代谢的各个步骤中，如转录和剪接。 解旋酶使用来自A1 'P水解的能量将双链核酸分离成 单股RNA解旋酶A对于小鼠的正常发育是必需的， 苍蝇和模式生物C.优雅其精确的细胞功能是 未知，但在体外实验与人类RHA同源物提供了一些 线索例如，体外转录需要人RHA CBP激活RNA聚合酶II，CBP是一种参与cAMP信号传导的蛋白质， 牢房CBP的突变导致人类疾病Rubinstein-Taybi综合征， 这与发育异常有关，如宽指， 智力迟钝。人RHA还负责基因组病毒RNA输出 从原子核中分离出来。为了确定RHA的精确细胞功能， 这项建议涉及对RHA- 1的生物化学和遗传学研究， C.优雅RNA结合、ATP酶和RNA解旋酶性质 将使用纯化的蛋白质和明确的RNA在体外研究RHA-1 印刷受体.在细胞中可能与RHA- 1结合的候选蛋白质将是 使用酵母双杂交系统进行测试。这些结合伙伴可能揭示了 RHA-1的细胞功能，它们可能会改变RHA-1的生化活性。 1.最后，RHA- 1和/或其结合伴侣功能降低的蠕虫 将产生以确定RHA-1的生理功能。的 最终目的是确定RHA-1的细胞功能，以获得更好的 理解转录和mRNA加工如何影响发育 包括人类在内的所有真核生物。

DESCRIPTION (provided by applicant): Organisms develop in precisely organized patterns that are controlled by complex cellular mechanisms. Many of these mechanisms are conserved throughout higher eukaryotes. Some proteins are responsible for directing the development of specific parts of the body (e.g. homeotic proteins). Others, such as RNA helicase A (RHA), are involved in basic cellular processes in all cells, and defects in these proteins produce developmental abnormalities throughout the organism. RNA helicases are involved in various steps of mRNA metabolism, such as transcription and splicing. Helicases use energy from A1'P hydrolysis to separate duplex nucleic acids into single strands. RNA helicase A is essential for proper development in mice, flies, and the model organism C. elegans. Its precise cellular function is unknown, but in vitro experiments with the human RHA homolog provide some clues. For example, human RHA is required for the in vitro transcription activation of RNA polymerase II by CBP, a protein involved in cAMP signaling in the cell. Mutations in CBP cause the human disease Rubinstein-Taybi syndrome, which is associated with developmental abnormalities such as broad digits and mental retardation. Human RHA is also responsible for genomic viral RNA export from the nucleus. In order to determine the precise cellular function of RHA, this proposal involves biochemical and genetic studies of RHA- 1, the RHA homolog in C. elegans. RNA binding, ATPase, and RNA helicase properties of RHA-1 will be studied in vitro using purified protein and well-defined RNA substrates. Candidate proteins that may bind to RHA- 1 in the cell will be tested using the yeast two-hybrid system. These binding partners may reveal the cellular function of RHA-1 and they may alter the biochemical activity of RHA- 1. Finally, worms with reduced function of RHA- 1 and/or its binding partners will be generated to determine the physiological function of RHA-1. The ultimate goal is to determine the cellular function of RHA-1 to obtain a better understanding of how transcription and mRNA processing affect the development in all eukaryotes, including humans.