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XB-ART-58836
PLoS Genet January 18, 2022; 18 (1): e1010012.
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Systematic mapping of rRNA 2''-O methylation during frog development and involvement of the methyltransferase Fibrillarin in eye and craniofacial development in Xenopus laevis.

Delhermite J , Tafforeau L , Sharma S , Marchand V , Wacheul L , Lattuca R , Desiderio S , Motorin Y , Bellefroid E , Lafontaine DLJ .


Abstract
Ribosomes are essential nanomachines responsible for protein production. Although ribosomes are present in every living cell, ribosome biogenesis dysfunction diseases, called ribosomopathies, impact particular tissues specifically. Here, we evaluate the importance of the box C/D snoRNA-associated ribosomal RNA methyltransferase fibrillarin (Fbl) in the early embryonic development of Xenopus laevis. We report that in developing embryos, the neural plate, neural crest cells (NCCs), and NCC derivatives are rich in fbl transcripts. Fbl knockdown leads to striking morphological defects affecting the eyes and craniofacial skeleton, due to lack of NCC survival caused by massive p53-dependent apoptosis. Fbl is required for efficient pre-rRNA processing and 18S rRNA production, which explains the early developmental defects. Using RiboMethSeq, we systematically reinvestigated ribosomal RNA 2''-O methylation in X. laevis, confirming all 89 previously mapped sites and identifying 15 novel putative positions in 18S and 28S rRNA. Twenty-three positions, including 10 of the new ones, were validated orthogonally by low dNTP primer extension. Bioinformatic screening of the X. laevis transcriptome revealed candidate box C/D snoRNAs for all methylated positions. Mapping of 2''-O methylation at six developmental stages in individual embryos indicated a trend towards reduced methylation at specific positions during development. We conclude that fibrillarin knockdown in early Xenopus embryos causes reduced production of functional ribosomal subunits, thus impairing NCC formation and migration.

PubMed ID: 35041640
Article link: PLoS Genet


Species referenced: Xenopus laevis
Genes referenced: ctrl dmrta2 fbl gapdh hes4 krt12.1 krt12.5 msx1 ncl nog nop58 npat npb pax2 pax6 prdm12 slc12a3 sox2 tp53 twist1 ubtf wnt8a
Antibodies: FLAG Ab1 Fbl Ab1 Tuba4a Ab3
Morpholinos: fbl MO1 ncl MO1 ubtf MO1

Phenotypes: Xla Wt + fbl MO (Fig. 2 C) [+]

Article Images: [+] show captions
References [+] :
Aittaleb, Structure and function of archaeal box C/D sRNP core proteins. 2003, Pubmed