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The Shwachman-Bodian-Diamond syndrome protein mediates translational activation of ribosomes in yeast

Abstract

The autosomal recessive disorder Shwachman-Diamond syndrome, characterized by bone marrow failure and leukemia predisposition, is caused by deficiency of the highly conserved Shwachman-Bodian-Diamond syndrome (SBDS) protein. Here, we identify the function of the yeast SBDS ortholog Sdo1, showing that it is critical for the release and recycling of the nucleolar shuttling factor Tif6 from pre-60S ribosomes, a key step in 60S maturation and translational activation of ribosomes. Using genome-wide synthetic genetic array mapping, we identified multiple TIF6 gain-of-function alleles that suppressed the pre-60S nuclear export defects and cytoplasmic mislocalization of Tif6 observed in sdo1Δ cells. Sdo1 appears to function within a pathway containing elongation factor–like 1, and together they control translational activation of ribosomes. Thus, our data link defective late 60S ribosomal subunit maturation to an inherited bone marrow failure syndrome associated with leukemia predisposition.

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Figure 1: TIF6 gain-of-function alleles can bypass the requirement for SDO1.
Figure 2: Translational impairment and aminoglycoside sensitivity of sdo1 mutant cells is rescued by TIF6 gain-of-function alleles.
Figure 3: Sdo1 is required for 60S ribosomal subunit biogenesis.
Figure 4: Tif6 suppressor proteins show reduced binding to 60S ribosomes.
Figure 5: Sdo1 cofractionates with cytoplasmic pre-60S ribosomes and does not shuttle in a Crm1-dependent manner.
Figure 6: SDS macrophages show redistribution of eIF6 protein.
Figure 7: Model for Sdo1 function in late 60S ribosomal subunit maturation.

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Acknowledgements

We thank E. Hurt (University of Heidelberg) for providing plasmids expressing Rpl25-eGFP, Rps2-eGFP and DsRed-Nop1; B. Trumpower (Dartmouth Medical School) for providing Rpl10 antisera; the Yeast Resource Center (http://www.yeastrc.org/pdr/) for providing strain TDY619; D. Tollervey for discussion of unpublished data; S. Munro (MRC Laboratory of Molecular Biology) for discussion and strains SEY6210 and BY4741; J. Kilmartin (MRC Laboratory of Molecular Biology) for discussion and plasmid pFA6-KanMX4; B. Andrews and J. Sale for critical reading of the manuscript and C. Hilcenko for advice on data analysis. We thank A. Johnson (University of Texas) for CRM1-T539C strains and M. Blundell (Great Ormond St. Hospital) for technical advice. This work was supported by grants from the UK Leukaemia Research Fund (A.J.W. and T.M.), the MRC (A.J.W. and C.C.W.), the Raymond and Beverly Sackler Fund (T.M. and C.C.W.), Shwachman-Diamond Support and the Leukemia and Lymphoma Society. C.B. was supported by grants from the National Cancer Institute of Canada, the Canadian Institute of Health Research, Genome Canada and Genome Ontario.

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Correspondence to Alan J Warren.

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Supplementary information

Supplementary Fig. 1

Sdo1 depletion results in underaccumulation of 60S ribosomes. (PDF 166 kb)

Supplementary Fig. 2

[3H-methyl]-methionine pulse-chase labeling analysis of pre-rRNA processing in Sdo1-depleted cells. (PDF 611 kb)

Supplementary Fig. 3

Yeast Sdo1 and human SBDS FYSH domains are functionally interchangeable in sdo1Δ cells. (PDF 82 kb)

Supplementary Table 1

Yeast strains. (PDF 44 kb)

Supplementary Table 2

Primers. (PDF 17 kb)

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Menne, T., Goyenechea, B., Sánchez-Puig, N. et al. The Shwachman-Bodian-Diamond syndrome protein mediates translational activation of ribosomes in yeast. Nat Genet 39, 486–495 (2007). https://doi.org/10.1038/ng1994

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