Nakayama Koh, Frew Ian J, Hagensen Mette, Skals Marianne, Habelhah Hasem, Bhoumik Anindita, Kadoya Takayuki, Erdjument-Bromage Hediye, Tempst Paul, Frappell Peter B, Bowtell David D, Ronai Ze'ev
Department of Oncological Sciences, Mount Sinai School of Medicine, New York, New York 10029, USA.
Cell. 2004 Jun 25;117(7):941-52. doi: 10.1016/j.cell.2004.06.001.
Hypoxia-inducible factor-1alpha (HIF1alpha) is a central regulator of the cellular response to hypoxia. Prolyl-hydroxylation of HIF1alpha by PHD enzymes is prerequisite for HIF1alpha degradation. Here, we demonstrate that the abundance of PHD1 and PHD3 are regulated via their targeting for proteasome-dependent degradation by the E3 ubiquitin ligases Siah1a/2, under hypoxia conditions. Siah2 null fibroblasts exhibit prolonged PHD3 half-life, resulting in lower levels of HIF1alpha expression during hypoxia. Significantly, hypoxia-induced HIF1alpha expression was completely inhibited in Siah1a/2 null cells, yet could be rescued upon inhibition of PHD3 by RNAi. Siah2 targeting of PHD3 for degradation increases upon exposure to even mild hypoxic conditions, which coincides with increased Siah2 transcription. Siah2 null mice subjected to hypoxia displayed an impaired hyperpneic respiratory response and reduced levels of hemoglobin. Thus, the control of PHD1/3 by Siah1a/2 constitutes another level of complexity in the regulation of HIF1alpha during hypoxia.
缺氧诱导因子-1α(HIF1α)是细胞对缺氧反应的核心调节因子。PHD酶对HIF1α进行脯氨酰羟化是HIF1α降解的前提条件。在此,我们证明在缺氧条件下,E3泛素连接酶Siah1a/2通过将PHD1和PHD3靶向蛋白酶体依赖性降解来调节它们的丰度。Siah2基因缺失的成纤维细胞表现出PHD3半衰期延长,导致缺氧期间HIF1α表达水平降低。值得注意的是,在Siah1a/2基因缺失的细胞中,缺氧诱导的HIF1α表达被完全抑制,但通过RNAi抑制PHD3后可恢复。暴露于甚至轻度缺氧条件下,Siah2将PHD3靶向降解的作用增强,这与Siah2转录增加相吻合。遭受缺氧的Siah2基因缺失小鼠表现出呼吸急促反应受损和血红蛋白水平降低。因此,Siah1a/2对PHD1/3的调控构成了缺氧期间HIF1α调控中另一个复杂层面。