Proteomic analysis on the leaves of TaBTF3 gene virus-induced silenced wheat plants may reveal its regulatory mechanism

Abstract

Basic transcription factor 3 (BTF3) is involved in the transcriptional initiation of RNA polymerase II and is also associated with apoptosis. In this study, virus-induced gene silencing of TaBTF3 caused severe viral symptoms in wheat seedlings, which then displayed stunted growth, reduced height, and decreased total fresh and dry weights. A proteomic approach was further used to identify the protein species showing differential abundance between the TaBTF3 virus-induced gene silenced wheat plants and the barley stripe mosaic virus-induced gene silencing green fluorescent protein transgenic wheat plants (control) with the objective of exploring its regulatory mechanism in higher plants. Using two-dimensional electrophoresis technologies, 59 protein spots showed significant changes, of which 54 were successfully identified by tandem mass spectrometry with matrix-assisted laser desorption/ionization-time of flight spectrometry. Analysis of protein abundance revealed that the differential protein species were associated with signal transduction, stress defense, photosynthesis, carbohydrate metabolism, and protein metabolism, and were mostly localized in both chloroplasts and mitochondria. Furthermore, the BTF3-responsive protein interaction network revealed 20 key protein species, most of which are regulated by abscisic acid, ethane, or oxidative stress. This suggested that changes of these protein species could be critical in the BTF3 pathway. Basic transcription factor 3 (BTF3), the β-subunit of NAC, has originally been identified as a basic transcription factor that is both involved in the transcriptional initiation of RNA polymerase II and associated with diverse biological functions. Reports on BTF3 mainly focus in animals, however, there has been limited molecular information about BTF3 in higher plants so far. In previous studies, we first isolated the TaBTF3 gene from common wheat (Triticum aestivum L.) and obtained silenced transgenic wheat seedlings using the VIGS method. In TaBTF3-silenced transgenic wheat plants, the structure of the wheat mesophyll cell was seriously damaged and transcripts of the chloroplast- and mitochondrial-encoded genes were significantly reduced. These results suggested that the TaBTF3 gene may be involved in regulating the growth and development of wheat seedlings. However, the induced or related genes by TaBTF3 have not been identified. The significance of this study is to first identify many protein species with the altered abundance between the TaBTF3 virus-induced silencing wheat plants and the BSMV-VIGS GFP transgenic wheat plants (control) using the proteomic approach. In addition, 20 of these identified protein species which might play critical roles in the BTF3 interaction network are identified using protein interaction network. These results help to further explore the molecular mechanism of BTF3 in higher plants.