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Proteomic profiling of a fast neutron induced soybean mutant unveiled regulators of increased seed protein content

ABSTRACT: Mutagenesis through fast neutrons (FN) radiation in soybean resulted 15% increase in seed protein content of a mutant. A comparative genomic hybridizations (CGHs) analyses confirmed that the mutant is lacking of 24 genes located at chromosome 5, 10 and 15. A proteomic profiling of the wild type and a FN mutant revealed 3,502 proteins, of which 206 protein exhibited increased abundance and 214 decreased abundance. Among the abundant proteins, basic 7S globulin increased in four-fold, followed by vacuolar-sorting receptor and protein transporters. The differentially expressed proteins were mapped to the global metabolic pathways. A higher enrichment in ribosomal, endoplasmic reticulum and protein export metabolic pathways were observed. We anticipate that the deletion of the sequence-specific DNA binding transcription factor along with other 23 genes might have a cascaded effect of protein synthesis, resulting in an increased amount of 7S globulin

INSTRUMENT(S): Orbitrap Fusion mass spectrometer Lumos

ORGANISM(S): Glycine Max (ncbitaxon:3847)

SUBMITTER: Savithiry Natarajan (Savi)

PROVIDER: MSV000084228 | MassIVE | Fri Aug 23 08:20:00 BST 2019

REPOSITORIES: MassIVE

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Project description:Intake of high-protein (HP) diets has increased over the last years, mainly due to their popularity for body weight control. Liver is the main organ handling ingested macronutrients and it is associated with the beginning of different pathologies. We aimed to deepen our knowledge on molecular pathways affected by long-term intake of an HP diet. We performed a transcriptome analysis on liver of rats chronically fed with a casein-rich HP diet and analyzed molecular parameters related to liver injury. Chronic increase in the dietary protein/carbohydrate ratio up-regulated processes related with amino acid uptake/metabolism and lipid synthesis, promoting a molecular environment indicative of hepatic triacylglycerol (TG) deposition. Moreover, changes in expression of genes involved in acidâbase maintenance and oxidative stress indicate alterations in the pH balance due to the high acid load of the diet, which has been linked to liver/health damage. Up-regulation of immune-related genes was also observed. In concordance with changes at gene expression level, we observed increased liver TG content and increased serum markers of hepatic injury/inflammation (aspartate transaminase, C-reactive protein and TNF-alpha). Moreover, the HP diet strongly increased hepatic mRNA and protein levels of HSP90, a marker of liver injury. Thus, we show for the first time that long-term consumption of an HP diet, resulting in a high acid load, results in a hepatic transcriptome signature reflecting increased TG deposition and increased signs of health risk (increased inflammation, alterations in the acidâbase equilibrium and oxidative stress). Persistence of this altered metabolic status could have unhealthy consequences. Wistar wildtype male rats, aged 8 weeks, received a low fat diet, high protein or high fat diet for 4 months. After sacrification, livers were dissected, and immediately snap frozen in liquid nitrogen. Total RNA was isolated, quantified and qualified, and subsequently used for global gene expression profiling using Agilent 4x44K microarrays.

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Proteomic profiling of a fast neutron induced soybean mutant unveiled regulators of increased seed protein content

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