Deubiquitinating enzyme mutagenesis screens identify a USP43-dependent HIF-1 transcriptional response.

IF 9.4 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
EMBO Journal Pub Date : 2024-09-01 Epub Date: 2024-07-15 DOI:10.1038/s44318-024-00166-6
Tekle Pauzaite, Niek Wit, Rachel V Seear, James A Nathan
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引用次数: 0

Abstract

The ubiquitination and proteasome-mediated degradation of Hypoxia Inducible Factors (HIFs) is central to metazoan oxygen-sensing, but the involvement of deubiquitinating enzymes (DUBs) in HIF signalling is less clear. Here, using a bespoke DUBs sgRNA library we conduct CRISPR/Cas9 mutagenesis screens to determine how DUBs are involved in HIF signalling. Alongside defining DUBs involved in HIF activation or suppression, we identify USP43 as a DUB required for efficient activation of a HIF response. USP43 is hypoxia regulated and selectively associates with the HIF-1α isoform, and while USP43 does not alter HIF-1α stability, it facilitates HIF-1 nuclear accumulation and binding to its target genes. Mechanistically, USP43 associates with 14-3-3 proteins in a hypoxia and phosphorylation dependent manner to increase the nuclear pool of HIF-1. Together, our results highlight the multifunctionality of DUBs, illustrating that they can provide important signalling functions alongside their catalytic roles.

去泛素化酶诱变筛选确定了一种依赖于 USP43 的 HIF-1 转录反应。
低氧诱导因子(HIFs)的泛素化和蛋白酶体介导的降解是后生动物氧传感的核心,但去泛素化酶(DUBs)参与 HIF 信号传导的情况还不太清楚。在这里,我们利用定制的 DUBs sgRNA 文库进行 CRISPR/Cas9 诱变筛选,以确定 DUBs 如何参与 HIF 信号传导。在确定参与 HIF 激活或抑制的 DUBs 的同时,我们还发现 USP43 是有效激活 HIF 响应所需的 DUB。USP43 受缺氧调节,并选择性地与 HIF-1α 异构体结合,虽然 USP43 不会改变 HIF-1α 的稳定性,但它能促进 HIF-1 的核积累并与其靶基因结合。从机理上讲,USP43 以缺氧和磷酸化依赖的方式与 14-3-3 蛋白结合,以增加 HIF-1 的核库。我们的研究结果凸显了 DUBs 的多功能性,说明它们在发挥催化作用的同时还能提供重要的信号功能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
EMBO Journal
EMBO Journal 生物-生化与分子生物学
CiteScore
18.90
自引率
0.90%
发文量
246
审稿时长
1.5 months
期刊介绍: The EMBO Journal has stood as EMBO's flagship publication since its inception in 1982. Renowned for its international reputation in quality and originality, the journal spans all facets of molecular biology. It serves as a platform for papers elucidating original research of broad general interest in molecular and cell biology, with a distinct focus on molecular mechanisms and physiological relevance. With a commitment to promoting articles reporting novel findings of broad biological significance, The EMBO Journal stands as a key contributor to advancing the field of molecular biology.
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