RNA-NetHOX · Decoding Hox specificity from mRNA processing networks
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2022-01-01 → 2023-12-31
- EU contribution
- €116,954
- Participants
- 1
- Scheme
- MSCA-IF
Lines connect the coordinator with its partners.
Results in brief
Decoding Hox specificity from mRNA processing networks
Eukaryotic gene expression is remarkable: from limited genetic material, it creates various proteomes and cell types. To do so, genes are transcribed into pre-mRNAs that undergo processing including splicing, the mechanism of exon/intron retention or excision, to produce mature mRNAs. A single pre-mRNA can be spliced in different ways thereby diversifying transcript isoforms and proteins. This mechanism termed alternative splicing contributes to the diversity of cell and tissue identities in complex organisms. Conversely, aberrant splicing leads to severe pathologies such as neuromuscular disorders and cancers. However, realising how such transcriptional and splicing programs are coordinated is still a challenge in Biology. Transcription factors (TFs) are the key players in gene expression by triggering precise spatial-temporal transcriptional programs. If most TFs act at the DNA regulatory layer, some TFs can bind RNA and modulate mRNA splicing. Yet, the mechanistic clues underlying TF function in alternative splicing remain elusive. Solving this issue will provide unique entry points to understand the mechanisms orchestrating cell and tissue diversity in animals and their aberrant regulation in diseases. To address this central issue in gene regulation, we use the Hox TF as a model based on our recently published work. Specifically, the research objectives were the following: -Decipher how interactions with splicing factors could impact Hox tissue-specific functions -Enlarge the cooperative role of mRNA processing regulators and HOX proteins As outcomes of the project, we have characterised the interaction between the Hox TF Ultrabithorax (Ubx) and splicing factors in the mesoderm of Drosophila embryos. We have generated a genetic toolkit for studying Ubx splicing function and the toolkit for assessing the Ubx/splicing factors function in vivo is in progress. We have determined the conservation of HOX/splicing factors interaction in humans and established a protocol to identify the interactome of these complexes. Capturing these complexes and characterising their function as outlook will contribute to a better understanding of the mechanisms governing cell and tissue diversity and their deregulation in diseases.
Data: CORDIS, © European Union
Project objective
Cell fate decisions is governed by the fine-tuned regulation of gene expression in all living organism. It is mostly realized by the cell-type specific assembly of regulatory networks consisting of Transcription Factors (TFs) and cofactors proteins. Thus, a multitude of combination between TF and cofactors will be set up to regulate the transcription in a precise spatiotemporal manner. The evolutionary conserved class of Hox TFs perfectly illustrates how a restricted number of TFs is able to promote diverse transcriptional programs. Hox proteins are involved in the specification of body forms and organs in animals. Despite this notorious architectural role, their operating mode remains controversial: they recognize similar DNA-binding sites in vitro, in sharp contrast with their specific functions in vivo. Thus, Hox proteins are likely acting with cofactors in vivo, and their identification was the core of my post-doctoral work. Notably, it revealed that mRNA-processing related proteins could constitute an important class of Hox cofactors. Thus, Hox specificity could also rely on mRNA-processing regulation, providing insightful molecular entry points to understand Hox function in development and disease. My research project aims at elucidating this novel facet of Hox activity. I propose two complementary aims developed in two model systems in order to decipher the molecular and functional impact of Hox-dependent mRNA processing regulation. I will decipher how interactions with splicing factors impact on the Hox functions in vivo in Drosophila embryos. Moreover, I will enlarge the cooperative role of HOX and splicing factors by performing large-scale interaction-screens of several human HOX and 2000 RNA-regulatory proteins in various normal and pathological cell contexts. Overall, my research program has the ambition to open novel perspectives on the role of TFs at the mRNA-regulatory level.
Original text from CORDIS.
Participants
- CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS · ParisCoordinatorFrance
Links
Data: CORDIS, © European Union