PNAS published a new paper of Pascal describing a new technique called HaloTag-NAPPA (Halo Tag nucleic acid programmable protein array).
PNAS published a new paper of Pascal describing a new technique called HaloTag-NAPPA (Halo Tag nucleic acid programmable protein array). Congratulation Pascal!
Mapping transcription factor interactome networks using HaloTag protein arrays
Yazaki J, Galli M, Kim AY, Nito K, Aleman F, Chang KN, Carvunis AR, Quan R,
Nguyen H, Song L, Alvarez JM, Huang SC, Chen H, Ramachandran N, Altmann S,
Gutiérrez RA, Hill DE, Schroeder JI, Chory J, LaBaer J, Vidal M, Braun P, Ecker
JR. Proc Natl Acad Sci U S A. 2016 Jun 29. pii: 201603229.
This array was used to query protein-protein interaction of certrain transcription factors and regulators.The resulting transcription factor interactome network contains thousands of novel interactions.
Pressemitteilung TUM:
http://www.tum.de/
http://www.tum.de/die-tum/aktuelles/pressemitteilungen/kurz/article/33223/
Abstract:
Protein microarrays enable investigation of diverse biochemical properties for thousands of proteins in a single experiment, an unparalleled capacity. Using a high-density system called HaloTag nucleic acid programmable protein array (HaloTag-NAPPA), we created high-density protein arrays comprising 12,000 Arabidopsis ORFs. We used these arrays to query protein-protein interactions for a set of 38 transcription factors and transcriptional regulators (TFs) that function in diverse plant hormone regulatory pathways. The resulting transcription factor interactome network, TF-NAPPA, contains thousands of novel interactions. Validation in a benchmarked in vitro pull-down assay revealed that a random subset of TF-NAPPA validated at the same rate of 64% as a positive reference set of literature-curated interactions. Moreover, using a bimolecular fluorescence complementation (BiFC) assay, we confirmed in planta several interactions of biological interest and determined the interaction localizations for seven pairs. The application of HaloTag-NAPPA technology to plant hormone signaling pathways allowed the identification of many novel transcription factor-protein interactions and led to the development of a proteome-wide plant hormone TF interactome network.