A novel ribosome-dimerization protein found in the hyperthermophilic archaeon Pyrococcus furiosus using ribosome- associated proteomics

Ribosome dimerization is one of the bacterial events that suppresses protein synthesis in the stationary phase. Protein factors responsible for ribosome dimerization in bacteria are well characterized, whereas no information is available for the corresponding factors in archaeal and eukaryotic cells. Here we describe a protein found among the ribosome-associated proteins which dimerizes the 30S ribosomal subunit of the archaeon Pyrococcus furiosus. The ribosome-associated proteins were prepared by high salt wash of crude ribosomes, and analyzed by nanoflow liquid chromatography-tandem mass spectrometry (nano LC-MS/MS). Of the detected proteins we focused on a protein (PF0560) whose Protein Score was the highest of all of the function-unknown proteins. PF0560 protein had a pronounced effect on the sedimentation pattern of the 30S ribosomal subunit; addition of this protein to isolated 30S subunit reduced the 30S fraction and increased the amount of the 50S fraction. This increase presumably corresponds to the dimer of the 30S subunit. The PF0560-dependent 30S-dimerization, was also observed by gel electrophoretic analysis. This effect was not observed in EDTA-treated 30S subunit, with protein-free 16S rRNA or with bacterial/eukaryotic ribosomal small subunits. Furthermore, PF0560 protein suppressed the formation of functional 70S ribosomes. These results suggest that PF0560 is a novel 30S dimerization factor, which might participate in regulation of archaeal translation. (c) 2022 Published by Elsevier Inc.

Automated summary

Ribosome dimerization is an event that suppresses protein synthesis in bacteria during the stationary phase. While the protein factors responsible for ribosome dimerization in bacteria are well characterized, little is known about the corresponding factors in archaeal and eukaryotic cells. In this study, a protein (PF0560) was identified that dimerizes the 30S ribosomal subunit in the archaeon Pyrococcus furiosus. This protein had a pronounced effect on the sedimentation pattern of the 30S subunit, indicating the formation of 30S dimers. Furthermore, the protein suppressed the formation of functional 70S ribosomes. These findings suggest that PF0560 is a novel dimerization factor involved in the regulation of translation in archaea.