NMR structure; mRNA splicing; translation regulation; IRES; splicing regulation; U1snRNP; RNP; protein-RNA recognition
Cieniková Zuzana, Jayne Sandrine, Damberger Fred Franz, Allain Frédéric Hai-Trieu, Maris Christophe (2015), Evidence for cooperative tandem binding of hnRNP C RRMs in mRNA processing., in RNA (New York, N.Y.)
, 21(11), 1931-42.
Afroz Tariq, Skrisovska Lenka, Belloc Eulàlia, Guillén-Boixet Jordina, Méndez Raúl, Allain Frédéric H-T (2014), A fly trap mechanism provides sequence-specific RNA recognition by CPEB proteins., in Genes & development
, 28(13), 1498-514.
Duss Olivier, Yulikov Maxim, Jeschke Gunnar, Allain Frédéric H-T (2014), EPR-aided approach for solution structure determination of large RNAs or protein-RNA complexes., in Nature communications
, 5, 3669-3669.
Duss Olivier, Michel Erich, Diarra dit Konté Nana, Schubert Mario, Allain Frédéric H-T (2014), Molecular basis for the wide range of affinity found in Csr/Rsm protein-RNA recognition., in Nucleic acids research
, 42(8), 5332-46.
Cieniková Zuzana, Damberger Fred F, Hall Jonathan, Allain Frédéric H-T, Maris Christophe (2014), Structural and mechanistic insights into poly(uridine) tract recognition by the hnRNP C RNA recognition motif., in Journal of the American Chemical Society
, 136(41), 14536-44.
Duss Olivier, Michel Erich, Yulikov Maxim, Schubert Mario, Jeschke Gunnar, Allain Frédéric H-T (2014), Structural basis of the non-coding RNA RsmZ acting as a protein sponge., in Nature
, 509(7502), 588-92.
Understanding the molecular mechanisms regulating gene expression at the posttranscriptional level is a major question in molecular biology, defects in these mechanisms being at the origin of many genetic diseases. This proposal focuses on two of these processes: pre-mRNA splicing regulation and translation regulation. Posttranscriptional gene regulation plays a major role in many diverse cellular processes such as embryonic development, cell differentiation and synapse activation. We focus here on aspects of translational and splicing regulation under the control of proteins binding the mRNA and pre-mRNA. At the mRNA 5’UTR, we are interested in how hnRNP proteins (PTB and UNR) control IRES-mediated translation initiation of viral and cellular mRNAs. At the mRNA 3’UTR, we are interested in how RNA binding proteins like Dnd1 regulate translation by counteracting translation repression by miRNA. In splicing regulation, we wish to understand how alternative-splicing factors like hnRNP A1 or C assemble upon pre-mRNA binding but also how factors like PTB or SR proteins interact with the splicing machinery and in particular with U1snRNP.We are proposing here to determine the NMR structures of several key protein-RNA complexes involved in translation and splicing regulation. First, we aim at understanding how PTB and Unr bind IRES to facilitate or repress translation initiation, second, how DND1 binds mRNA 3’end to counteract miRNAs, third how multimers of hnRNPA1 or hnRNP C assemble on the pre-mRNA and finally how U1snRNP assembles, recognizes the 5’ splice-site and interacts with various alternative-splicing factors (PTB, SR proteins, TIA-1). To solve the smaller protein-RNA complexes, we will apply the same NOE-based approach that we have used successfully with others ranging in size from 15 to 30 kDa. For the larger complexes, we will start by determining the structures of stable smaller sub-complexes and subsequently use them as building blocks to reassemble the largest complex using RDCs, EPR, PRE and/or SAXS data.