Deeba Noreen Baig

Distortion of the normal function of synaptic cell adhesion molecules by genetic variants as a risk for autism spectrum disorders

Synaptic cell adhesion molecules (SCAMs) are a functional category of cell adhesion molecules that connect pre- and postsynapses by the protein-protein interaction via their extracellular cell adhesion domains. Countless numbers of common genetic variants and rare mutations in SCAMs have been identified in the patients with autism spectrum disorders (ASDs). Among these, NRXN and NLGN family proteins cooperatively function at synaptic terminals both of which genes are strongly implicated as risk genes for ASDs. Knock-in mice carrying a single rare point mutation of NLGN3 (NLGN3 R451C) discovered in the patients with ASDs display a deficit in social interaction and an enhancement of spatial learning and memory ability reminiscent of the clinical phenotype of ASDs. NLGN4 knockout (KO) and NRXN2α KO mice also show a deficit in sociability as well as some specific neuropsychiatric behaviors. In this review, we selected NRXNs/NLGNs, CNTNAP2/CNTNAP4, CNTN4, ITGB3, and KIRREL3 as strong ASD risk genes based on SFARI score and summarize the protein structures, functions at synapses, representative discoveries in human genetic studies, and phenotypes of the mutant model mice in light of the pathophysiology of ASDs.

Secondary Metabolites Production and Plant Growth Promotion by Pseudomonas chlororaphis and P. aurantiaca Strains Isolated from Cactus, Cotton, and Para Grass

Fluorescent pseudomonads have been isolated from halophytes, mesophytes, and xerophytes of Pakistan. Among these, eight isolates, GS-1, GS-3, GS-4, GS-6, GS-7, FS-2 (cactus), ARS-38 (cotton), and RP-4 (para grass), showed antifungal activity and were selected for detailed study. Based on biochemical tests and 16S rRNA gene sequences, these were identified as strains of P. chlororaphis subsp. chlororaphis and aurantiaca. Secondary metabolites of these strains were analyzed by LC-MS. Phenazine-1-carboxylic acid (PCA), 2-hydroxy-phenazine, Cyclic Lipopeptide (white line-inducing principle (WLIP)), and lahorenoic acid A were detected in variable amounts in these strains. P. aurantiaca PB-St2 was used as a reference as it is known for the production of these compounds. The phzO and PCA genes were amplified to assure that production of these compounds is not an artifact. Indole acetic acid production was confirmed and quantified by HPLC. HCN and siderophore production by all strains was observed by plate assays. These strains did not solubilize phosphate, but five strains were positive for zinc solubilization. Wheat seedlings were inoculated with these strains to observe their effect on plant growth. P. aurantiaca strains PB-St2 and GS-6 and P. chlororaphis RP-4 significantly increased both root and shoot dry weights, as compared with uninoculated plants. However, P. aurantiaca strains FS-2 and ARS-38 significantly increased root and shoot dry weights, respectively. All strains except PB-St2 and ARS-38 significantly increased the root length. This is the first report of the isolation of P. aurantiaca from cotton and cactus, P. chlororaphis from para grass, WLIP and lahorenoic acid A production by P. chlororaphis, and zinc solubilization by P. chlororaphis and P. aurantiaca.

Contribution of Zinc Solubilizing Bacteria in Growth Promotion and Zinc Content of Wheat

Zinc is an imperative micronutrient required for optimum plant growth. Zinc solubilizing bacteria are potential alternatives for zinc supplementation and convert applied inorganic zinc to available forms. This study was conducted to screen zinc solubilizing rhizobacteria isolated from wheat and sugarcane, and to analyze their effect on wheat growth and development. Fourteen exo-polysaccharides producing bacterial isolates of wheat were identified and characterized biochemically as well as on the basis of 16S rRNA gene sequences. Along these, 10 identified sugarcane isolates were also screened for zinc solubilizing ability on five different insoluble zinc sources. Out of 24, five strains, i.e., EPS 1 (Pseudomonas fragi), EPS 6 (Pantoea dispersa), EPS 13 (Pantoea agglomerans), PBS 2 (E. cloacae) and LHRW1 (Rhizobium sp.) were selected (based on their zinc solubilizing and PGP activities) for pot scale plant experiments. ZnCO3 was used as zinc source and wheat seedlings were inoculated with these five strains, individually, to assess their effect on plant growth and development. The effect on plants was analyzed based on growth parameters and quantifying zinc content of shoot, root and grains using atomic absorption spectroscopy. Plant experiment was performed in two sets. For first set of plant experiments (harvested after 1 month), maximum shoot and root dry weights and shoot lengths were noted for the plants inoculated with Rhizobium sp. (LHRW1) while E. cloacae (PBS 2) increased both shoot and root lengths. Highest zinc content was found in shoots of E. cloacae (PBS 2) and in roots of P. agglomerans (EPS 13) followed by zinc supplemented control. For second set of plant experiment, when plants were harvested after three months, Pantoea dispersa (EPS 6), P. agglomerans (EPS 13) and E. cloacae (PBS 2) significantly increased shoot dry weights. However, significant increase in root dry weights and maximum zinc content was recorded for Pseudomonas fragi (EPS 1) inoculated plants, isolated from wheat rhizosphere. While maximum zinc content for roots was quantified in the control plants indicating the plants inability to transport zinc to grains, supporting accelerated bioavailability of zinc to plant grains with zinc solubilizing rhizobacteria.

Distribution analysis of profilin isoforms at transcript resolution with mRNA-seq and secondary structure in various organs of Rattus norvegicus.

Profilin (Pfn) is an actin binding protein, ubiquitously found in mammals and is essential for the actin polymerization in cells. In brain, it plays a pivotal role in neurogenesis and synapse formation by interacting with various proteins. Four Pfn isoforms have been identified in mammals. This study presents the identification and transcriptional expression of various Pfn isoforms (Pfn1, Pfn2, Pfn3 and Pfn4) in brain, heart, kidney, liver, and muscle and testis of Rattus norvegicus. Organs have been classified into groups based on some similarities. Group I includes brain and testis, Group II includes skeletal muscle and heart, while Group III includes kidney and liver. Pfn1 has been identified in all groups, Pfn2 and Pfn3 have been identified in group I, group III and in one organ (skeletal muscle) of group II. To the best of the authors knowledge, no report of Pfn1 and Pfn2 presence in testis, Pfn3 in brain, liver and skeletal muscle, Pfn4 in kidney and skeletal muscle exists to date. Transcriptional expression showed variations among expression level of different Pfn isoforms in various organs with respect to the control gene GADPH. We hypothesize that this could be attributed to profilin isoform specific mRNA structure and corresponding motifs, which generally contribute to similar or varied decay rates, cellular localization, post transcriptional regulation pattern and ligand binding.

Digital dissection of arsenate reductase enzyme from an arsenic hyperccumulating fern Pteris vittata

Action of arsenate reductase is crucial for the survival of an organism in arsenic polluted area. Pteris vittata, also known as Chinese ladder brake, was the first identified arsenic hyperaccumulating fern with the capability to convert [As(V)] to arsenite [As(III)]. This study aims at sequence analysis of the most important protein of the arsenic reduction mechanism in this specie. Phosphorylation potential of the protein along with possible interplay of phosphorylation with O-β-GlcNAcylation was predicted using neural network based webservers. Secondary and tertiary structure of arsenate reductase was then analysed. Active site region of the protein comprised a rhodanese-like domain. Cursory dynamics simulation revealed that folds remained conserved in the rhodanese main but variations were observed in the structure in other regions. This information sheds light on the various characteristics of the protein and may be useful to enzymologists working on the improvement of its traits for arsenic reduction.

Deciphering brain-specific transcriptomic expression of Ezr , Rad and Msn genes in the development of Mus musculus

Ezrin, Radixin and Moesin (ERM) are critical membranous component involved in cross‐linking of actin filaments. Moesin (Msn) is recognized as a pivotal protein involved in regulation of cell signalling events associated with the maintenance of epithelial integrity, actin organization and polarity. Radixin (Rad) is known to cell‐to‐cell adherens junction as a barbed end‐capping protein whereas ezrin (Ezr) is recognized at cell adhesion, motility, apoptosis and phagocytosis. The current study for the first time reports the transcriptional and RNA secondary structural variations among brain‐specific ERM genes. Firstly, we analyzed brain‐specific transcriptomic expression in selected embryonic and postnatal developmental stages (E10.5, E14.5, E18.5, P0.5, P3.5, P5.5, P10.5 and P20.5) of Mus musculus. Among designated developmental stages, Ezr has highest fold difference in early embryonic and postnatal stages (E10.5, P0.5 and P5.5). Rad showed a similar pattern of high expression especially at embryonic stages (E10.5 and E18.5) and postnatal (P0.5 and P5.5), however, Msn exhibited non‐significant fold differences in comparison to controls leading to its curial role in development. Furthermore, computational prediction of ERM coding mRNA transcripts, reveals compact and less dynamic Msn secondary structure and pseudoknots configurations, in contrast to Ezr and Rad. Conclusively, transcriptomic levels are greatly associated with compact base pairing organization of its secondary structures. These findings open a new domain to understand the occurrence of ERM‐specific cytoskeleton proteins during developmental stages.

Prediction of post translation modifications at the contact site between Anaplasma phagocytophilum and human host during autophagosome induction using a bioinformatic approach.

Autophagy is crucial for maintaining physiological homeostasis, but its role in infectious diseases is not yet adequately understood. The binding of Anaplasma translocated substrate-1 (ATS1) to the human Beclin1 (BECN1) protein is responsible for the modulation of autophagy pathway. ATS1-BECN1 is a novel type of interaction that facilitates Anaplasma phagocytophilum proliferation, leading to intracellular infection via autophagosome induction and segregation from the lysosome. Currently, there is no report of post translational modifications (PTMs) of BECN1 or cross-talk required for ATS-BECN1 complex formation. Prediction/modeling of the cross-talk between phosphorylation and other PTMs (O-β-glycosylation, sumoylation, methylation and palmitoylation) has been attempted in this study, which might be responsible for regulating function after the interaction of ATS1 with BECN1. PTMs were predicted computationally and mapped onto the interface of the docked ATS1-BECN1 complex. Results show that BECN1 phosphorylation at five residues (Thr91, Ser93, Ser96, Thr141 and Ser234), the interplay with O-β-glycosylation at three sites (Thr91, Ser93 and Ser96) with ATS1 may be crucial for attachment and, hence, infection. No other PTM site at the BECN1 interface was predicted to associate with ATS1. These findings may have significant clinical implications for understanding the etiology of Anaplasma infection and for therapeutic studies.