biostudies-literatureLin JFHunan Provincial Natural Science Foundation of ChinaPostdoctoral Science Foundation of ChinaGuangdong Basic and Applied Basic Research FoundationNational Natural Science Foundation of ChinaThe National Key R&D Program of ChinaNational Natural Science Foundation of China (National Science Foundation of China)Young Talents Program of Sun Yat-sen University Cancer Center263https://www.ebi.ac.uk/biostudies/studies/S-EPMC11696352biostudies-literatureUnknown16(1)Metabolic enzymes perform moonlighting functions during tumor progression, including the modulation of chemoresistance. However, the underlying mechanisms of these functions remain elusive. Here, utilizing a metabolic clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 knockout library screen, we observe that the loss of glutamate-cysteine ligase modifier subunit (GCLM), a rate-limiting enzyme in glutathione biosynthesis, noticeably increases the sensitivity of colorectal cancer (CRC) cells to platinum-based chemotherapy. Mechanistically, we unveil a noncanonical mechanism through which nuclear GCLM competitively interacts with NF-kappa-B (NF-κB)-repressing factor (NKRF), to promote NF-κB activity and facilitate chemoresistance. In response to platinum drug treatment, GNature communicationsNucleus-translocated GCLM promotes chemoresistance in colorectal cancer through a moonlighting function.PMC116963528230330682373376Liang ZBLin JFChen DLZeng ZLHuang RZCao FHan YLiu ZXYu KSheng HLiao KXu RHLi TJu HQLi SSGao SMo HYfalseNucleus-translocated GCLM promotes chemoresistance in colorectal cancer through a moonlighting function.Metabolic enzymes perform moonlighting functions during tumor progression, including the modulation of chemoresistance. However, the underlying mechanisms of these functions remain elusive. Here, utilizing a metabolic clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 knockout library screen, we observe that the loss of glutamate-cysteine ligase modifier subunit (GCLM), a rate-limiting enzyme in glutathione biosynthesis, noticeably increases the sensitivity of colorectal cancer (CRC) cells to platinum-based chemotherapy. Mechanistically, we unveil a noncanonical mechanism through which nuclear GCLM competitively interacts with NF-kappa-B (NF-κB)-repressing factor (NKRF), to promote NF-κB activity and facilitate chemoresistance. In response to platinum drug treatment, G2025-01-01T00:00:00Z2025 Jan2026-06-02T02:04:18.85Z2025-04-05T22:50:01.759ZS-EPMC116963523974710110.1038/s41467-024-55568-1