TY  - JOUR
AU  - Rabany, Ofri
AU  - Ben Dror, Sivan
AU  - Arafat, Maram
AU  - Aharoni Levitanus, Hadar
AU  - Halperin, Yudit
AU  - Marchand, Virginie
AU  - Romanovski, Nikolai
AU  - Ussishkin, Noga
AU  - Livneh Golany, Maayan
AU  - Reches, Adi
AU  - Wexler, Judith
AU  - Mayorek, Nina
AU  - Monderer-Rothkoff, Galya
AU  - Shifman, Sagiv
AU  - Mâmmer Bouhou, Widad
AU  - VanInsberghe, Michael
AU  - Pauli, Cornelius
AU  - Müller-Tidow, Carsten
AU  - Karmi, Ola
AU  - Livneh, Yoav
AU  - van Oudenaarden, Alexander
AU  - Motorin, Yuri
AU  - Nachmani, Daphna
TI  - Dynamic rRNA Methylation Regulates Translation in the Hematopoietic System and is Essential for Stem Cell Fitness.
JO  - Blood
VL  - nn
SN  - 0006-4971
CY  - Washington, DC
PB  - American Society of Hematology
M1  - DKFZ-2025-02287
SP  - nn
PY  - 2025
N1  - epub
AB  - Self-renewal and differentiation are at the basis of hematopoiesis. While it is known that tight regulation of translation is vital for hematopoietic stem cells' (HSCs) biology, the mechanisms underlying translation regulation across the hematopoietic system remain obscure. Here we reveal a novel mechanism of translation regulation in the hematopoietic hierarchy, which is mediated by ribosomal RNA (rRNA) methylation dynamics. Using ultra-low input ribosome-profiling, we characterized cell-type-specific translation capacity during erythroid differentiation. We found that translation efficiency changes progressively with differentiation and can distinguish between discrete cell populations as well as to define differentiation trajectories. To reveal the underlying mechanism, we performed comprehensive mapping of the most abundant rRNA modification - 2'-O-methyl (2'OMe). We found that, like translation efficiency, 2'OMe dynamics followed a distinct trajectory during erythroid differentiation.Genetic perturbation of individual 2'OMe sites demonstrated their distinct roles in modulating proliferation and differentiation. By combining CRISPR screening, molecular and functional analyses, we identified a specific methylation site, 28S-Gm4588, which is progressively lost during differentiation, as a key regulator of HSC self-renewal. We showed that low methylation at this site led to translational skewing, mediated mainly by codon frequency, which promoted differentiation. Functionally, HSCs with diminished 28S-Gm4588 methylation exhibited impaired self-renewal capacity ex-vivo, and loss of fitness in-vivo in bone marrow transplantations.Extending our findings beyond the hematopoietic system, we also found distinct dynamics of 2'OMe profiles during differentiation of non-hematopoietic stem cells. Our findings reveal rRNA methylation dynamics as a general mechanism for cell-type-specific translation, required for cell function and differentiation.
LB  - PUB:(DE-HGF)16
C6  - pmid:41191530
DO  - DOI:10.1182/blood.2024028300
UR  - https://inrepo02.dkfz.de/record/305655
ER  -