title: Presence of Not5 and ubiquitinated Rps7A in polysome fractions depends upon the Not4 E3 ligase.
abstract: In this study, we determine that Saccharomyces cerevisiae Not4 E3 ligase ubiquitinates Rps7A in vivo and in vitro, but not its paralogue, Rps7B. Ubiquitinated Rps7A is detectable only in 80S and polysomes, but not in free 40S fractions. A different role of the Rps7 paralogues in vivo is supported by the observation that the deletion of Rps7A but not Rps7B is sensitive to translational inhibitors and leads to an accumulation of aggregated proteins. An important accumulation of aggregated proteins that include ribosomal proteins and ribosome-associated chaperones is also observed in cells lacking Not4. A contribution of Not4 to ribosomal function extending beyond Rps7A ubiquitination is supported by the observation that the deletion of Not4 displays a synthetic slow growth phenotype when combined with the deletion of either one of the two Rps7 paralogues. Not4 is detectable in polysome fractions, as are other subunits of the Ccr4-Not complex such as Not5. The optimal presence of Not5 in polysomes is dependent upon Not4 and the deletion of Not5 leads to a dramatic reduction of polysomes. These results lead us to suggest that Not4 contributes to normal polysome levels and is important for cellular protein solubility maybe in part by ubiquitination of Rps7A.
authors.affiliation: Department of Microbiology and Molecular Medicine, University of Geneva, Faculty of Medicine, 1 rue Michel Servet, 1211 Geneva 4, Switzerland. olesya.panasenko@unige.ch
authors.forename: Olesya O
authors.initials: OO
authors.lastname: Panasenko
authors.forename: Martine A
authors.initials: MA
authors.lastname: Collart
mesh_terms: D000595:Amino Acid Sequence; D017353:Gene Deletion; D008969:Molecular Sequence Data; D011132:Polyribosomes / Q000378:metabolism*; D012097:Repressor Proteins; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism; D054875:Ubiquitination
chemical_list: C111175:NOT5 protein, S cerevisiae; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C086923:MOT2 protein, S cerevisiae; D044767:Ubiquitin-Protein Ligases