u4atac regulates cilium biogenesis through splicing of the minor intron of tmem107l and rfx7b in zebrafish developing brain
Bi-allelic variants of RNU4ATAC, transcribed into the minor spliceosome component U4atac snRNA, are associated to variable severity of microcephaly, growth retardation, skeletal dysplasia and immunodeficiency as main features. Previous studies highlighted the dramatic effect of U4atac deficiency on splicing of U12-type introns, which represent less than 1% of all introns in the human genome. More recently, our team evidenced a link between U4atac and the primary cilium/centrosome complex through the identification of patients carrying RNU4ATAC bi-allelic variants and exhibiting an atypical Joubert syndrome, a well-known ciliopathy. Yet, the underlying mechanisms remain elusive. Here, we further explored the link of RNU4ATAC to primary cilium and aimed at identifying ciliary U12-type intron containing genes that contribute to the brain abnormalities seen in patients. For that, we performed a transcriptomic analysis of heads of our morpholino oligonucleotide (MO)-mediated u4atac zebrafish model. Through the combined analysis of the generated dataset with those obtained from RNU4ATAC patient cells, we identified two candidate genes: TMEM107, coding for a structural protein of the cilium transition zone, and RFX7, encoding a transcription factor involved in primary cilium formation. By conducting complementary genetic approaches in zebrafish model, we showed that both gene orthologues, tmem107l and rfx7b, functionally interact with u4atac and are required for correct brain development. Altogether, our findings establish TMEM107 and RFX7 as key components of the molecular pathway linking U4atac dysfunction to ciliary defects and impaired brain development, providing new physiopathological insights and therapeutic perspectives for RNU4ATAC-related disorders.