Proteomic and Biochemical Analyses Show a Functional Network of Proteins Involved in Antioxidant Defense of the Arabidopsis anp2anp3 Double Mutant

Abstract

Disentanglement of functional complexity associated with plant mitogen-activated protein kinase (MAPK) signaling has benefited from transcriptomic, proteomic, phosphoproteomic, and genetic studies. Published transcriptomic analysis of a double homozygous recessive anp2anp3 mutant of two MAPK kinase kinase (MAPKKK) genes called Arabidopsis thaliana Homologues of Nucleus- and Phragmoplast-localized Kinase 2 (ANP2) and 3 (ANP3) showed the upregulation of stress-related genes. In this study, a comparative proteomic analysis of anp2anp3 mutant against its respective Wassilevskaja ecotype (Ws) wild type background is provided. Such differential proteomic analysis revealed overabundance of core enzymes such as FeSOD1, MnSOD, DHAR1, and FeSOD1-associated regulatory protein CPN20, which are involved in the detoxification of reactive oxygen species in the anp2anp3 mutant. The proteomic results were validated at the level of single protein abundance by Western blot analyses and by quantitative biochemical determination of antioxidant enzymatic activities. Finally, the functional network of proteins involved in antioxidant defense in the anp2anp3 mutant was physiologically linked with the increased resistance of mutant seedlings against paraquat treatment.

The Tomato Mitogen-Activated Protein Kinase SlMPK1 Is as a Negative Regulator of the High-Temperature Stress Response

“…As a first step, we analyzed proteomic data and found that several SlMPK1-mediated homologous proteins also were observed in the Arabidopsis MAPKKK double mutant anp2anp3 (Takáč et al, 2014). A comparative proteomic analysis of anp2anp3 revealed an overabundance of core enzymes such as SOD, DHAR1, and the FeSOD1-associated regulatory protein CPN20, which ensure favorable cellular redox conditions as well as accelerated defense against the overproduction of ROS (Takáč et al, 2014). In this study, many proteins controlling the oxidation-reduction process also were discovered (Supplemental Table S6).…”

Comprehensive analysis of transcriptional regulation and functional genes induced by alkaline salt stress in roots of two switchgrass (Panicum virgatum L.) genotypes

“…The CDPK and MAPK pathways are important signalling pathways in plant responses to salt stress [55]. The MAPK cascade is an important regulator of antioxidant defence [56]. CAMS/CMLS, CIPKs and CDPKs, which function as Ca 2+ sensors, are involved in many growth, development and stress-induced signal transduction pathways [57,58].…”

Advantages and limitations of shot-gun proteomic analyses on Arabidopsis plants with altered MAPK signaling

“…Shot-gun proteomic analyses on mutant and transgenic plants with genetically modified MAPKs and their upstream activators such as MAP2Ks and MAP3Ks provide a very useful proteome readout and quite unique information because they might not be fully consistent with transcriptomic data from the same plant lines (e.g., Krysan et al, 2002 cf. Takáč et al, 2014 ). Few studies used shot-gun proteomic approaches to better explain stress-related and developmental phenotypes in Arabidopsis mutants and transgenic lines with altered MAPKs expression.…”

“…Modern OMICS technologies encompassing transcriptomic (Krysan et al, 2002 ; Frei dit Frey et al, 2014 ), metabolomic (Lassowskat et al, 2014 ), proteomic (Miles et al, 2009 ; Conroy et al, 2013 ; Lassowskat et al, 2014 ; Takáč et al, 2014 ) and phosphoproteomic (Hoehenwarter et al, 2013 ; Lassowskat et al, 2014 ) approaches have been employed to gain functional information on plant signaling. The main challenges of advanced (phospho)proteomic studies are: reliable identification of all signaling constituents of a particular MAPK pathway as well as identification and validation of respective target proteins.…”