Computational Analysis of the Tripartite Interaction of Phasins (PhaP4 and 5)-Sigma Factor (σ24)-DNA of Azospirillum brasilense Sp7.

Azospirillum brasilense Sp7 produces PHB, which is covered by granule-associated proteins (GAPs). Phasins are the main GAPs. Previous studies have shown phasins can regulate PHB synthesis. When A. brasilense grows under stress conditions, it uses sigma factors to transcribe genes for survival. One of these factors is the σ24 factor. This study determined the possible interaction between phasins and the σ24 factor or phasin-σ24 factor complex and DNA. Three-dimensional structures of phasins and σ24 factor structures were predicted using the I-TASSER and SWISS-Model servers, respectively. Subsequently, a molecular docking between phasins and the σ24 factor was performed using the ClusPro 2.0 server, followed by molecular docking between protein complexes and DNA using the HDOCK server. Evaluation of the types of ligand-receptor interactions was performed using the BIOVIA Discovery Visualizer for three-dimensional diagrams, as well as the LigPlot server to obtain bi-dimensional diagrams. The results showed the phasins (Pha4Abs7 or Pha5Abs7)-σ24 factor complex was bound near the -35 box of the promoter region of the phaC gene. However, in the individual interaction of PhaP5Abs7 and the σ24 factor, with DNA, both proteins were bound to the -35 box. This did not occur with PhaP4Abs7, which was bound to the -10 box. This change could affect the transcription level of the phaC gene and possibly affect PHB synthesis.

Development and validation of a multivariable genotype-informed gestational diabetes prediction algorithm for clinical use in the Mexican population: insights into susceptibility mechanisms.

INTRODUCTION: Gestational diabetes mellitus (GDM) is underdiagnosed in Mexico. Early GDM risk stratification through prediction modeling is expected to improve preventative care. We developed a GDM risk assessment model that integrates both genetic and clinical variables. RESEARCH DESIGN AND METHODS: Data from pregnant Mexican women enrolled in the 'Cuido mi Embarazo' (CME) cohort were used for development (107 cases, 469 controls) and data from the 'Mónica Pretelini Sáenz' Maternal Perinatal Hospital (HMPMPS) cohort were used for external validation (32 cases, 199 controls). A 2-hour oral glucose tolerance test (OGTT) with 75 g glucose performed at 24-28 gestational weeks was used to diagnose GDM. A total of 114 single-nucleotide polymorphisms (SNPs) with reported predictive power were selected for evaluation. Blood samples collected during the OGTT were used for SNP analysis. The CME cohort was randomly divided into training (70% of the cohort) and testing datasets (30% of the cohort). The training dataset was divided into 10 groups, 9 to build the predictive model and 1 for validation. The model was further validated using the testing dataset and the HMPMPS cohort. RESULTS: Nineteen attributes (14 SNPs and 5 clinical variables) were significantly associated with the outcome; 11 SNPs and 4 clinical variables were included in the GDM prediction regression model and applied to the training dataset. The algorithm was highly predictive, with an area under the curve (AUC) of 0.7507, 79% sensitivity, and 71% specificity and adequately powered to discriminate between cases and controls. On further validation, the training dataset and HMPMPS cohort had AUCs of 0.8256 and 0.8001, respectively. CONCLUSIONS: We developed a predictive model using both genetic and clinical factors to identify Mexican women at risk of developing GDM. These findings may contribute to a greater understanding of metabolic functions that underlie elevated GDM risk and support personalized patient recommendations.

Phasin PhaP1 is involved in polyhydroxybutyrate granules morphology and in controlling early biopolymer accumulation in Azospirillum brasilense Sp7.

Phasins are amphiphilic proteins involved in the regulation of the number and size of polyhydroxybutyrate (PHB) granules. The plant growth promoting bacterium Azospirillum brasilense Sp7 accumulates high quantities of bioplastic PHB as carbon and energy source. By analyzing the genome, we identified six genes that code for proteins with a Phasin_2 domain. To understand the role of A. brasilense Sp7 PhaP1 (PhaP1Abs) on PHB synthesis, the phaP1 gene (AMK58_RS17065) was deleted. The morphology of the PHB granules was analyzed by transmission electron microscopy (TEM) and the PHB produced was quantified under three different C:N ratios in cultures subjected to null or low-oxygen transfer. The results showed that PhaP1Abs is involved in PHB granules morphology and in controlling early biopolymer accumulation. Using RT-PCR it was found that phasin genes, except phaP4, are transcribed in accordance with the C:N ratio used for the growth of A. brasilense. phaP1, phaP2 and phaP3 genes were able to respond to the growth conditions tested. This study reports the first analysis of a phasin protein in A. brasilense Sp7.