Coorey Namal V, Tollestrup Isaac, Bircham Peter W, Sheridan Jeffrey P, Evans Gary B, Schramm Vern L, Atkinson Paul H, Munkacsi Andrew B
School of Biological Sciences, Victoria University of Wellington, Wellington 6012, New Zealand.
Ferrier Research Institute, Victoria University of Wellington, Wellington 6012, New Zealand.
Dis Model Mech. 2025 Jun 1;18(6). doi: 10.1242/dmm.052173. Epub 2025 Jun 30.
Methylthioadenosine-DADMe immucillin-A (MTDIA) is a transition-state analog that potently inhibits the human protein 5'-methylthioadenosine phosphorylase (MTAP) at picomolar concentrations and elicits anti-tumor activity against lung, prostate, colon, cervical, head and neck, and triple-negative breast cancers in cell and animal models. The anti-cancer mechanisms of MTDIA involve elevated methylthioadenosine levels but are not fully understood. The yeast protein MEU1 is functionally equivalent to human MTAP. To gain further understanding, we performed chemical genetic analyses via gene deletion and GFP-tagged protein libraries in yeast that express a member of the human equilibrative nucleoside transporter (ENT) family to permit MTDIA uptake. Genomic and proteomic analyses identified genes and proteins critical to MTDIA bioactivity. Network analysis of these genes and proteins revealed an important link to ribosomal function, which was confirmed by observing reduced levels of ribosomal subunit proteins. Network analysis also implicated autophagy, which was confirmed by analyzing intracellular trafficking of GFP-Atg8 and Phloxine B viability. In yeast, a comparable effect occurred after deletion of MEU1, indicating a single target for MTDIA in yeast. Overall, our yeast model reveals specific components of the ribosome as well as induction of autophagy as integral mechanisms that mediate the bioactivity of MTDIA.
甲硫基腺苷 - DADMe免疫菌素 - A（MTDIA）是一种过渡态类似物，在皮摩尔浓度下能有效抑制人蛋白5'-甲硫基腺苷磷酸化酶（MTAP），并在细胞和动物模型中对肺癌、前列腺癌、结肠癌、宫颈癌、头颈癌和三阴性乳腺癌产生抗肿瘤活性。MTDIA的抗癌机制涉及甲硫基腺苷水平升高，但尚未完全了解。酵母蛋白MEU1在功能上等同于人类MTAP。为了进一步了解，我们通过基因缺失和在表达人类平衡核苷转运体（ENT）家族成员以允许MTDIA摄取的酵母中的绿色荧光蛋白（GFP）标记蛋白文库进行了化学遗传学分析。基因组和蛋白质组分析确定了对MTDIA生物活性至关重要的基因和蛋白质。对这些基因和蛋白质的网络分析揭示了与核糖体功能的重要联系，这通过观察核糖体亚基蛋白水平降低得到证实。网络分析还涉及自噬，这通过分析GFP - Atg8的细胞内运输和焰红B活力得到证实。在酵母中，缺失MEU1后也出现了类似的效果，表明MTDIA在酵母中有单一靶点。总体而言，我们的酵母模型揭示了核糖体的特定组成部分以及自噬的诱导是介导MTDIA生物活性的整体机制。
