Interaction proteomics of the HMGA chromatin architectural factors.
The high mobility group A (HMGA) chromatin architectural transcription factors are a group of proteins involved in development and neoplastic transformation. They take part in an articulated interaction network, both with DNA and other nuclear proteins, organizing multimolecular complexes at chromatin level. Here, we report the development of a novel in vitro strategy for the identification of HMGA molecular partners based on the combination of an RP-HPLC prefractionation procedure, 2-DE gels, blot-overlay and MS. To demonstrate that our approach could be a reliable screening method we confirmed a representative number of interactions in vitro by GST pull-down and far-Western and in vivo by co-affinity purification. This approach allowed us to enlarge the HMGA molecular network confirming their involvement also in non-transcriptional-related processes such as RNA processing and DNA repair.
Sgarra
Riccardo
R
Dipartimento di Scienze della Vita, Università degli Studi di Trieste, Italy.
Furlan
Cristina
C
Zammitti
Salvina
S
Lo Sardo
Alessandra
A
Maurizio
Elisa
E
Di Bernardo
Julie
J
Giancotti
Vincenzo
V
Manfioletti
Guidalberto
G
D000818:Animals; D002471:Cell Transformation, Neoplastic; D002843:Chromatin / Q000378:metabolism*; D002853:Chromatography, Liquid; D004260:DNA Repair; D015180:Electrophoresis, Gel, Two-Dimensional; D053263:Gene Regulatory Networks; D006609:High Mobility Group Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping; D040901:Proteomics / Q000379:methods*; D012323:RNA Processing, Post-Transcriptional; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry
D002843:Chromatin; D006609:High Mobility Group Proteins; D011994:Recombinant Proteins