Chemical proteomic profiling of UTP-binding proteins in human cells.

Nucleotide-binding proteins play important roles in a variety of biological processes. While ATP- and GTP-binding proteins have been well studied, the systematical identification of UTP-interacting proteins remains under investigated. Here, we developed a chemical proteomic strategy using a biotinylated UTP affinity probe coupled with liquid chromatography tandem mass spectrometry (LC-MS/MS) method to enrich, identify and quantify UTP-binding proteins at the entire proteome scale. By performing labeling reactions with high vs low concentrations of UTP probe (100 and 10 µM) or with the UTP probe in the presence of free UTP in stable isotope labeling by amino acids in cell culture (SILAC) experiments, we identified more than 70 potential UTP-binding proteins which are involved in multiple cellular processes, such as translational elongation and protein folding. We also validated the UTP-binding capability of the cytoskeletal protein ACTB by using cellular thermal shift assay (CETSA). Together, we performed a high-throughput chemical proteomics-based analysis to identify, for the first time, UTP-binding proteins in human proteome, which should be applicable for the identification and quantification of UTP-binding proteins in other organisms.

MicroRNA-4317 predicts the prognosis of breast cancer and inhibits tumor cell proliferation, migration, and invasion.

Previous researches have indicated that miR-4317 was aberrantly expressed in several tumors. However, the potential role of miR-4317 in breast cancer is still unclear. The aim of this study was to investigate the potential role of miR-4317 in breast cancer. The relative expression levels of miR-4317 were detected in breast cancer tissues and cell lines using qRT-PCR analysis. The Kaplan-Meier survival curve and multivariate Cox regression analyses were used to investigate the prognostic significance of miR-4317 in breast cancer. CCK-8 and Transwell assays were performed to evaluate the effects of miR-4317 on cell proliferation, migration, and invasion. The results showed that miR-4317 expression was decreased in breast cancer tissues and cell lines. Downregulation of miR-4317 was significantly associated with lymph node metastasis, TNM stage, and poor prognosis. Overexpression of miR-4317 inhibited proliferation, migration, and invasion of breast cancer cells, while downregulation of miR-4317 exhibited the opposite effects. MYD88 may be a direct target of miR-4317. The results suggest miR-4317 may play a tumor suppressor role in breast cancer and inhibit proliferation, migration, and invasion of breast cancer cells by targeting MYD88. The findings provide novel evidence of miR-4317 as a potential prognostic biomarker and therapeutic target for breast cancer.