| Literature DB >> 26230714 |
Tiago Antonio de Souza, Adriana Santos Soprano, Nayara Patricia Vieira de Lira, Alexandre José Christino Quaresma, Bianca Alves Pauletti, Adriana Franco Paes Leme, Celso Eduardo Benedetti.
Abstract
Entities:
Year: 2015 PMID: 26230714 PMCID: PMC4521785 DOI: 10.1371/journal.pone.0134818
Source DB: PubMed Journal: PLoS One ISSN: 1932-6203 Impact factor: 3.240
Fig 1Protein-protein interactions between PthAs and citrus nuclear proteins.
(A) Citrus preys fused to yeast GAL4-AD (GAL4AD-prey) or control plasmid (GAL4AD) were moved into yeast cells carrying one of the four PthA variants fused to GAL4-BD domain as shown in the diagram (1 to 4, respectively). Yeast double-transformants were grown on SC-Trp-Leu-His-Ade in the presence of 5 mM of 3AT. None of prey fusions transactivated the reporter genes when co-transformed with empty bait vector (5). The PthA baits also did not transactivate the reporter genes when co-transformed with the empty prey vector in the same growth conditions (GAL4AD). (B) Western blot detection of eluted fractions from GST pulldown assays using the purified 6xHis-PthAs 1–4 as prey and immobilized GST or GST-fusion proteins as baits. Arrows indicate bands corresponding to the expected size for the GST-fusion proteins CsHMG (~45 kDa), CsTRAX (~55 kDa), CsSMC (~45 kDa), CsRRPMP1 (~50 kDa), CsRRMP2 (~46 kDa), CsPABP1 (~53 kDa) and CsVIP2 (~85 kDa) detected by the GST anti-serum. PthA proteins (~116–122 kDa) were detected using the anti-PthA serum. Recombinant PthAs 3 and 4 were added as references in the first lanes of the gels shown in the middle and right panels, respectively.