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ZNF692 organizes a hub for ribosome maturation enhancing translation in rapidly proliferating cells

ABSTRACT: Human cancer cell lines (DLD1 wt or ZNF692 KO, and for IP-proteomics HCT116 transfected with GFP, GFP-ZNF692 and deltaNolsZNF692). 788570, HCT116 transfected with GFP; 788571, HCT116 transfected with GFP-ZNF692; 788572, HCT116 transfected with deltaNolsZNF692; 937612, control 40S subunit from DLD1 cells; 937613, control 60S subunit from DLD1 cells; 937614, control 80S monosome fraction from DLD1 cells; 937615, KO ZNF692 40S subunit from DLD1 cells; 937616, KO ZNF692 60S subunit from DLD1 cells; 937617, KO ZNF692 80S monosome fraction from DLD1 cells; 943031, control 40S subunit from DLD1 cells; 943032, control 60S subunit from DLD1 cells; 943033, control 80S monosome fraction from DLD1 cells; 943034, KO ZNF692 40S subunit from DLD1 cells; 943035, KO ZNF692 60S subunit from DLD1 cells; 943036, KO ZNF692 80S monosome fraction from DLD1 cells

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Maralice Conacci-Sorrell

PROVIDER: MSV000089271 | MassIVE | Mon Apr 18 12:41:00 BST 2022

REPOSITORIES: MassIVE

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Project description:The REIL proteins are required for late ribosomal biogenesis and accumulation of the 60S large ribosome subunit in mature leaves of Arabidopsis thaliana upon acclimation to low temperature. To validate these functions in roots, we conducted a multi-level system analysis targeted at understanding defects and compensations responses of reil mutants before acclimation to low temperature and following temperature shift. Hydroponic root tissue enabled analysis of eukaryotic ribosome complexes with negligible interference of organelle ribosomes. Hydroponic cultivation attenuated the growth defect of reil mutants at low temperature and provided new insights into the primary functions of Arabidopsis REIL proteins. Arabidopsis tightly controls the balance of non-translating 40S and 60S subunits. Reil mutants initially deplete both non-translating subunits upon shift to 10°C and subsequently replenish these pools slowly. Reil mutations compensate the 60S biosynthesis defect by increased baseline levels of non-translating 40S and 60S subunits and depletion of a likely non-translating, KCl-sensitive 80S sub-fraction in the cold. We infer that Arabidopsis buffers fluctuating translation demands following temperature cues by activating non-translating ribosome fractions before de novo synthesis meets temperature-induced demands. Reil1 reil2 double mutants accumulate 43S-preinitiation complexes and pre-60S-maturation complexes and affect the paralog composition of non-translating ribosome fractions. With few exceptions, e.g. RPL3B and RPL24C, these changes were not under transcriptional control. Our study suggests requirement of de novo synthesis of eukaryotic ribosomes for long-term cold acclimation. Double mutant analysis indicates feedback control of REIL-mediated 60S maturation on NUC2 and eIF3C2 transcription and implies functions of two so far non-described proteins in late plant ribosome biogenesis. We propose that Arabidopsis requires biosynthesis of specialized ribosomes for successful cold acclimation.

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ZNF692 organizes a hub for ribosome maturation enhancing translation in rapidly proliferating cells

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