The uL10 protein, a component of the ribosomal P-stalk, is released from the ribosome in nucleolar stress

Biochim Biophys Acta Mol Cell Res. 2018 Jan;1865(1):34-47. doi: 10.1016/j.bbamcr.2017.10.002. Epub 2017 Oct 3.

Abstract

The ribosomal uL10 protein, formerly known as P0, is an essential element of the ribosomal GTPase-associated center responsible for the interplay with translational factors during various stages of protein synthesis. In eukaryotic cells, uL10 binds two P1/P2 protein heterodimers to form a pentameric P-stalk, described as uL10-(P1-P2)2, which represents the functional form of these proteins on translating ribosomes. Unlike most ribosomal proteins, which are incorporated into pre-ribosomal particles during early steps of ribosome biogenesis in the nucleus, P-stalk proteins are attached to the 60S subunit in the cytoplasm. Although the primary role of the P-stalk is related to the process of translation, other extraribosomal functions of its constituents have been proposed, especially for the uL10 protein; however, the list of its activities beyond the ribosome is still an open question. Here, by the combination of biochemical and advanced fluorescence microscopy techniques, we demonstrate that upon nucleolar stress induction the uL10 protein accumulates in the cytoplasm of mammalian cells as a free, ribosome-unbound protein. Importantly, using a novel approach, FRAP-AC (FRAP after photoConversion), we have shown that the ribosome-free pool of uL10 represents a population of proteins released from pre-existing ribosomes. Taken together, our data indicate that the presence of uL10 on the ribosomes is affected in stressed cells, thus it might be considered as a regulatory element responding to environmental fluctuations.

Keywords: FRAP; Nucleolar stress; Photoconvertible fluorescent protein; Ribosomal protein; Ribosome.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arabidopsis Proteins / metabolism*
  • Cell Nucleolus / metabolism*
  • Chromosomal Proteins, Non-Histone / metabolism*
  • DNA-Binding Proteins / metabolism*
  • HeLa Cells
  • Humans
  • Mice
  • NIH 3T3 Cells
  • Protein Binding
  • Ribosomal Proteins / metabolism*
  • Ribosomes / metabolism*
  • Stress, Physiological / physiology*

Substances

  • Arabidopsis Proteins
  • BAF60 protein, Arabidopsis
  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • RPLP0 protein, human
  • Ribosomal Proteins