Functional mapping of the interaction between TDP-43 and hnRNP A2 in vivo.
Nuclear factor TDP-43 has been reported to play multiple roles in transcription, pre-mRNA splicing, mRNA stability and mRNA transport. From a structural point of view, TDP-43 is a member of the hnRNP protein family whose structure includes two RRM domains flanked by the N-terminus and C-terminal regions. Like many members of this family, the C-terminal region can interact with cellular factors and thus serve to modulate its function. Previously, we have described that TDP-43 binds to several members of the hnRNP A/B family through this region. In this work, we set up a coupled minigene/siRNA cellular system that allows us to obtain in vivo data to address the functional significance of TDP-43-recruited hnRNP complex formation. Using this method, we have finely mapped the interaction between TDP-43 and the hnRNP A2 protein to the region comprised between amino acid residues 321 and 366. Our results provide novel details of protein-protein interactions in splicing regulation. In addition, we provide further insight on TDP-43 functional properties, particularly the lack of effects, as seen with our assays, of the disease-associated mutations that fall within the TDP-43 321-366 region: Q331K, M337V and G348C.
D'Ambrogio
Andrea
A
International Centre for Genetic Engineering and Biotechnology (ICGEB), 34012 Trieste, Italy.
Buratti
Emanuele
E
Stuani
Cristiana
C
Guarnaccia
Corrado
C
Romano
Maurizio
M
Ayala
Youhna M
YM
Baralle
Francisco E
FE
D000595:Amino Acid Sequence; D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D010455:Peptides / Q000494:pharmacology; D025941:Protein Interaction Mapping; D034622:RNA Interference; D012326:RNA Splicing; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid
D004268:DNA-Binding Proteins; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D010455:Peptides; C439768:hnRNP A2; D019005:Cystic Fibrosis Transmembrane Conductance Regulator