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Christiane Branlant

Research Focus

Our lab has a long expertise on RNA and RNP structure-function analysis, especially, on spliceosomal UsnRNAs and UsnRNPs. This includes the study of post-transcriptional modifications in RNA and their biogenesis.

We are using HIV-1 RNA alternative splicing as a model to study the effect of pre-mRNA structure on alternative splicing. We developed an in cellulo system to investigate the effects of mutations in HIV-1 RNA on the relative utilisation of all splicing sites.

We demonstrated the involvement of RNA secondary structure on the interplay of splicing activators and inhibitors at two highly regulated acceptor sites of HIV-1 RNA (A3 and A7). This led to a new concept for 3' splice site regulation. We developed protocols for purification and characterisation of RNP complexes, formed at splicing sites, and are extending the analysis to human transcripts subjected to alternative splicing.

Publications

  1. Dreumont, N., Hardy, S., Behm-Ansmant, I., Kister, L., Branlant, C., Stévenin, J., Bourgeois, C.F. (2009). Antagonistic factors control the unproductive splicing of SC35 terminal intron. Nucleic Acids Research 2009 Dec 3. [Epub ahead of print]
  2. Saliou, J.M., Bourgeois, C.F., Mena, L.A., Ropers, D., Jacquenet, S., Marchand, V., Stévenin, J., Branlant, C. (2009). Role of RNA structure and protein factors in the control of HIV-1 splicing. Frontiers in Bioscience 14, 2714-29.
  3. Muller, S., Leclerc, F., Behm-Ansmant, I., Fourmann, J.B., Charpentier, B., Branlant, C. (2008). Combined in silico and experimental identification of the Pyrococcus abyssi H/ACA sRNAs and their target sites in ribosomal RNAs. Nucleic Acids Research 36(8), 2459-75.
  4. Boulon, S., Marmier-Gourrier, N., Pradet-Balade, B., Wurth, L., Verheggen, C., Jády, B.E., Rothé, B., Pescia, C., Robert, M.C., Kiss, T., Bardoni, B., Krol, A., Branlant, C., Allmang, C., Bertrand, E., Charpentier, B. (2008). The Hsp90 chaperone controls the biogenesis of L7Ae RNPs through conserved machinery. Journal of Cell Biology 180(3), 579-95.
  5. Piazzon, N., Rage, F., Schlotter, F., Moine, H., Branlant, C., Massenet, S. (2008). In vitro and in cellulo evidences for association of the survival of motor neuron complex with the fragile X mental retardation protein. Journal of Biological Chemistry 283(9), 5598-610.
  6. Bakkour, N., Lin, Y.L., Maire, S., Ayadi, L., Mahuteau-Betzer, F., Nguyen, C.H., Mettling, C., Portales, P., Grierson, D., Chabot, B., Jeanteur, P., Branlant, C., Corbeau, P., Tazi, J. (2007). Small-molecule inhibition of HIV pre-mRNA splicing as a novel antiretroviral therapy to overcome drug resistance. PLoS Pathogens 3(10), 1530-9.
  7. Muller, S., Fourmann, J.B., Loegler, C., Charpentier, B., Branlant, C. (2007). Identification of determinants in the protein partners aCBF5 and aNOP10 necessary for the tRNA:Psi55-synthase and RNA-guided RNA:Psi-synthase activities. Nucleic Acids Research 35(16), 5610-24.
  8. Behm-Ansmant, I., Branlant, C., Motorin, Y. (2007). The Saccharomyces cerevisiae Pus2 protein encoded by YGL063w ORF is a mitochondrial tRNA:Psi27/28-synthase. RNA 13(10), 1641-7.
  9. Charpentier, B., Fourmann, J.B., Branlant, C. (2007). Reconstitution of archaeal H/ACA sRNPs and test of their activity. Methods in Enzymology 425, 389-405.
  10. Muller, S., Charpentier, B., Branlant, C., Leclerc, F. (2007). A dedicated computational approach for the identification of archaeal H/ACA sRNAs. Methods in Enzymology 425, 355-87. PMID: 17673091 [PubMed - indexed for MEDLINE]
  11. Manival X., Charron C., Fourmann J. B.,. Godard F., Charpentier B. and Branlant C. (2006). Crystal structure determination and site-directed mutagenesis of the Pyrococcus abyssi aCBF5-aNOP10 complex reveal crucial roles of the C-terminal domains of both proteins in H/ACA sRNP activity. Nucleic Acids Research 34, 826-839.
  12. Ropers D., Ayadi-Behn-Mena L., Gattoni R., Jacquenet S., Damier L., Branlant C. and Stévenin J. (2004). Differential effects of the SR proteins 9G8, SC35, ASF/SF2 and SRp40 on the utilization of the A1 to A5 splicing sites of HIV-1 RNA. Journal of Biological Chemistry 279, 29963-29973.
  13. Mougin A., Gottschalk A., Fabrizio P., Lührmann R. and Branlant C. (2002). Direct probing of RNA structure and RNA-protein interactions in purified HeLa cell's and yeast spliceosomal U4/U6.U5 tri-snRNP particles. Journal of Molecular Biology 317, 631-649.
  14. Marchand V., Méreau A., Jacquenet S., Thomas D., Mougin A., Gattoni R., Stévenin J. C. Branlant C. (2002). A Janus Splicing Regulatory Element Modulates HIV-1 tat and rev mRNA Production by Coordination of hnRNP A1 Cooperative Binding. Journal of Molecular Biology 323, 629-652.
  15. Watkins, N., Ségault, V., Charpentier, B., Nottrott, S., Fabrizio, P., Bachi, A., Wilm, M., Rosbash, M., Branlant, C. and Lührmann, R. (2000). A common core RNP structure shared between the small nucleolar box C/D RNPs and the spliceosomal U4 snRNP. Cell 103, 457-466.

Key lab techniques: RNP purification using tagged RNAs, RNA and RNP structure analysis in solution, by X-ray cristallography and by in silico modelisation, in vitro and in cellulo experiments for splicing analysis.

Key lab reagents: Constructs for the study of HIV-1 RNA splicing.

Lab contact: Christiane Branlant: christiane.branlant@maem.uhp-nancy.fr