O'Connell et al. 2019 (PRJNA509230)

General Details

Title Mammalian Hbs1L deficiency causes Pelota depletion and is associated with a unique phenotype
Organism
Number of Samples 10
Release Date 2018/12/10 00:00
Sequencing Types
Protocol Details

Study Links

Repository Details

SRA SRP173240
ENA SRP173240
GEO
BioProject PRJNA509230

Publication

Title
Authors O'Connell AE,Gerashchenko MV,O'Donohue MF,Rosen SM,Huntzinger E,Gleeson D,Galli A,Ryder E,Cao S,Murphy Q,Kazerounian S,Morton SU,Schmitz-Abe K,Gladyshev VN,Gleizes PE,Séraphin B,Agrawal PB
Journal PLoS genetics
Publication Date 2019 Feb
Abstract Hbs1 has been established as a central component of the cell's translational quality control pathways in both yeast and prokaryotic models; however, the functional characteristics of its human ortholog (Hbs1L) have not been well-defined. We recently reported a novel human phenotype resulting from a mutation in the critical coding region of the HBS1L gene characterized by facial dysmorphism, severe growth restriction, axial hypotonia, global developmental delay and retinal pigmentary deposits. Here we further characterize downstream effects of the human HBS1L mutation. HBS1L has three transcripts in humans, and RT-PCR demonstrated reduced mRNA levels corresponding with transcripts V1 and V2 whereas V3 expression was unchanged. Western blot analyses revealed Hbs1L protein was absent in the patient cells. Additionally, polysome profiling revealed an abnormal aggregation of 80S monosomes in patient cells under baseline conditions. RNA and ribosomal sequencing demonstrated an increased translation efficiency of ribosomal RNA in Hbs1L-deficient fibroblasts, suggesting that there may be a compensatory increase in ribosome translation to accommodate the increased 80S monosome levels. This enhanced translation was accompanied by upregulation of mTOR and 4-EBP protein expression, suggesting an mTOR-dependent phenomenon. Furthermore, lack of Hbs1L caused depletion of Pelota protein in both patient cells and mouse tissues, while PELO mRNA levels were unaffected. Inhibition of proteasomal function partially restored Pelota expression in human Hbs1L-deficient cells. We also describe a mouse model harboring a knockdown mutation in the murine Hbs1l gene that shared several of the phenotypic elements observed in the Hbs1L-deficient human including facial dysmorphism, growth restriction and retinal deposits. The Hbs1lKO mice similarly demonstrate diminished Pelota levels that were rescued by proteasome inhibition.
PMC PMC6373978
PMID 30707697
DOI
Run Accession Study Accession Scientific Name Cell Line Library Type Treatment GWIPS-viz Trips-Viz Reads BAM BigWig (F) BigWig (R)
SRR8310196 PRJNA509230 Homo sapiens fibroblast Ribo-Seq Cycloheximide
SRR8310197 PRJNA509230 Homo sapiens fibroblast Ribo-Seq Cycloheximide
SRR8310198 PRJNA509230 Homo sapiens fibroblast Ribo-Seq Cycloheximide
SRR8310199 PRJNA509230 Homo sapiens fibroblast Ribo-Seq Cycloheximide
SRR8310200 PRJNA509230 Homo sapiens fibroblast Ribo-Seq Cycloheximide
SRR8310201 PRJNA509230 Homo sapiens fibroblast Ribo-Seq Cycloheximide
SRR8310202 PRJNA509230 Homo sapiens fibroblast Ribo-Seq Cycloheximide
SRR8310203 PRJNA509230 Homo sapiens fibroblast Ribo-Seq Cycloheximide
SRR8310204 PRJNA509230 Homo sapiens fibroblast Ribo-Seq Cycloheximide
SRR8310205 PRJNA509230 Homo sapiens fibroblast Ribo-Seq Cycloheximide
Run Accession Study Accession Scientific Name Cell Line Library Type Treatment GWIPS-viz Trips-Viz Reads BAM BigWig (F) BigWig (R)

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