Kothari KS, Dansana PK, Giri J, Tyagi AK.
Front Plant Sci. 2016 Jul 19;7:1057. doi: 10.3389/fpls.2016.01057. eCollection 2016.
Abstract
Stress associated proteins (SAPs) are the A20/AN1 zinc-finger containing proteins which can regulate the stress signaling in plants. The rice SAP protein, OsSAP1 has been shown to confer abiotic stress tolerance to plants, when overexpressed, by modulating the expression of endogenous stress-related genes. To further understand the mechanism of OsSAP1-mediated stress signaling, OsSAP1 interacting proteins were identified using yeast two-hybrid analysis. Two novel proteins, aminotransferase (OsAMTR1) and a SCP/TAPS or pathogenesis-related 1 class of protein (OsSCP) were found to interact with OsSAP1. The genes encoding OsAMTR1 and OsSCP were stress-responsive and showed higher expression upon abiotic stress treatments. The role of OsAMTR1 and OsSCP under stress was analyzed by overexpressing them constitutively in Arabidopsis and responses of transgenic plants were assessed under salt and water-deficit stress. The OsAMTR1 and OsSCP overexpressing plants showed higher seed germination, root growth and fresh weight than wild-type plants under stress conditions. Overexpression of OsAMTR1 and OsSCP affected the expression of many known stress-responsive genes which were not affected by the overexpression of OsSAP1. Moreover, the transcript levels of OsSCP and OsAMTR1 were also unaffected by the overexpression of OsSAP1. Hence, it was concluded that OsSAP1 regulates the stress responsive signaling by interacting with these proteins which further regulate the downstream stress responsive gene expression.
See http://www.ncbi.nlm.nih.gov/pubmed/27486471
FIGURE 1. Identification of proteins interacting with OsSAP1. (A) The DNA binding domain (BD) was fused to different deletions of OsSAP1 to prepare bait constructs. (B) Yeast cells co-transformed with bait constructs shown in (A) and AD clones (SCP-AD, and AMTR1-AD) were plated on SD/-Leu/-Trp (SD/-LT) and SD/-Ade/-His/-Leu/-Trp (SD/-AHLT) media to test the interaction. Empty BD vector was co-transformed with empty AD clones to serve as negative control. (C) Quantification of β-galactosidase activity in the transformed yeast cells using o-nitrophenyl-β-galactopyranoside (ONPG) as a substrate. Negative controls: Empty AD transformed with SAP1:BD. The histogram represents mean ± SE of three biological replicates.
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