Inositol Signaling Section, Laboratory of Signal Transduction, 111 T.W. Alexander Drive, Research Triangle Park, NC 27709, USA.
J Cell Sci. 2013 Jan 15;126(Pt 2):437-44. doi: 10.1242/jcs.110031. Epub 2012 Nov 30.
Fundamental to the life and destiny of every cell is the regulation of protein synthesis through ribosome biogenesis, which begins in the nucleolus with the production of ribosomal RNA (rRNA). Nucleolar organization is a highly dynamic and tightly regulated process; the structural factors that direct nucleolar assembly and disassembly are just as important in controlling rRNA synthesis as are the catalytic activities that synthesize rRNA. Here, we report that a signaling enzyme, inositol 1,3,4,5,6-pentakisphosphate 2-kinase (IP5K) is also a structural component in the nucleolus. We demonstrate that IP5K has functionally significant interactions with three proteins that regulate rRNA synthesis: protein kinase CK2, TCOF1 and upstream-binding-factor (UBF). Through molecular modeling and mutagenic studies, we identified an Arg-Lys-Lys tripeptide located on the surface of IP5K that mediates its association with UBF. Nucleolar IP5K spatial dynamics were sensitive to experimental procedures (serum starvation or addition of actinomycin D) that inhibited rRNA production. We show that IP5K makes stoichiometrically sensitive contributions to the architecture of the nucleoli in intact cells, thereby influencing the degree of rRNA synthesis. Our study adds significantly to the biological significance of IP5K; previously, it was the kinase activity of this protein that had attracted attention. Our demonstration that IP5K 'moonlights' as a molecular scaffold offers an unexpected new example of how the biological sophistication of higher organisms can arise from gene products acquiring multiple functions, rather than by an increase in gene number.
核糖体生物发生是调节蛋白质合成的基础,它始于核仁中 rRNA(核糖体 RNA)的产生。核仁组织是一个高度动态和严格调控的过程;指导核仁组装和拆卸的结构因子在控制 rRNA 合成方面与催化 rRNA 合成的活性同样重要。在这里,我们报告一种信号酶,即肌醇 1,3,4,5,6-五磷酸 2-激酶(IP5K)也是核仁中的结构成分。我们证明 IP5K 与三种调节 rRNA 合成的蛋白质具有功能上重要的相互作用:蛋白激酶 CK2、TCOF1 和上游结合因子(UBF)。通过分子建模和突变研究,我们确定了 IP5K 表面上的一个 Arg-Lys-Lys 三肽,该三肽介导其与 UBF 的结合。核仁中 IP5K 的空间动力学对实验程序(血清饥饿或放线菌素 D 加药)敏感,这些程序抑制了 rRNA 的产生。我们表明,IP5K 对完整细胞中核仁的结构做出了化学计量敏感性的贡献,从而影响 rRNA 的合成程度。我们的研究极大地增加了 IP5K 的生物学意义;以前,正是该蛋白质的激酶活性引起了人们的关注。我们证明 IP5K“兼职”作为分子支架,为高等生物的生物学复杂性如何源于获得多种功能的基因产物而不是通过增加基因数量提供了一个意想不到的新例子。