the Interaction Proteome of Ribosomal 40S Components PNO1 and NOB1 Using TurboID Proximity Labeling Technology

Abstract

Background The ribosome assembly factors PNO1 and NOB1 play crucial roles in the maturation of the 40S ribosomal small subunit. TurboID is an efficient biotin ligase that can biotinylate proteins in proximity to the target protein and is widely used to study complex biological processes within cells.Here, we utilized this technology to investigate the complex interaction network of PNO1 and NOB1 within cells. Results Through immunofluorescence experiments, we found that PNO1 and NOB1 have different localizations within cells. By identifying the proximal proteins biotinylated by PNO1-TurboID and NOB1-TurboID, we discovered 871 proteins interacting with PNO1 and 1044 proteins interacting with NOB1, with 663 proteins overlapping. These results suggest that PNO1 and NOB1 are extensively involved in various biological processes within the cell. Furthermore, we found that PNO1 and NOB1 interact with translation-related proteins EIF4B and EIF4G2, indicating that they may participate in the mRNA translation process. Conclusion The interaction proteome results of PNO1 and NOB1 suggest that ribosome assembly factors are not only involved in ribosome biogenesis but also couple with multiple biological processes within the cell, such as mRNA translation. This provides a foundation for understanding the complex biological processes within the cell.