Bhargava P.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research
Proteomics | Year: 2010
Brain is the most complex and least understood organ of the body. Recent research suggests that epigenetics of the brain may be behind the complex functions of this master organ. Yeast, the simplest eukaryote, had been the model for studying the complex physiology of higher eukaryotes, including humans. Current depth in understanding of mechanisms of gene regulation has been possible mainly because of the knowledge acquired by epigenetic studies on yeast while the research on the biochemistry and physiology of the brain has been tremendously benefitted by proteomic studies. The independent advances of research in both these fields are now converging. As the current understanding of epigenetics can be applied to understand the mysteries of normal brain function as well as various diseases, modern proteomic approaches can help find new therapeutic targets. © 2009 Wiley-VCH Verlag GmbH & Co.
Rajender S.,Central Drug Research Institute Council of Scientific and Industrial Research |
Pooja S.,Central Drug Research Institute Council of Scientific and Industrial Research |
Gupta N.J.,Institute of Reproductive Medicine |
Chakrabarty B.,Institute of Reproductive Medicine |
And 2 more authors.
Journal of Andrology | Year: 2011
End-organ resistance to androgens, called androgen insensitivity syndrome (AIS), is a rare disorder. The most common cause of AIS is mutations(s) in the androgen receptor (AR) gene; however, a significant number of these mutations have not been functionally analyzed. In the present study, we analyzed a case of complete AIS for mutations in the AR gene. Sequencing of the entire coding region of the AR gene revealed a 2650G.A mutation (mRNA sequence reference) in exon 4 of the gene, resulting in replacement of glycine with glutamate at codon 708 in the ligand-binding domain of the AR protein. The mutation was absent in 200 normal male individuals analyzed to look at its occurrence in general population. In vitro androgen-binding and transactivation assays showed that the mutation resulted in approximately 65% loss of ligand binding and almost complete loss of transactivation function. Complete AIS in this individual was due to a G708E substitution in the AR protein.
Matharu N.K.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research |
Hussain T.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research |
Sankaranarayanan R.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research |
Mishra R.K.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research
Journal of Molecular Biology | Year: 2010
Polycomb group (PcG) and trithorax group (trxG) proteins are chromatin-mediated regulators of a number of developmentally important genes including the homeotic genes. In Drosophila melanogaster, one of the trxG members, Trithorax like (Trl), encodes the essential multifunctional DNA binding protein called GAGA factor (GAF). While most of the PcG and trxG genes are conserved from flies to humans, a Trl-GAF homologue has been conspicuously missing in vertebrates. Here, we report the first identification of c- Krox/. Th-POK as the vertebrate homologue of GAF on the basis of sequence similarity and comparative structural analysis. The in silico structural analysis of the zinc finger region showed preferential interaction of vertebrate GAF with GAGA sites similar to that of fly GAF. We also show by cross-immunoreactivity studies that both fly and vertebrate GAFs are highly conserved and share a high degree of structural similarity. Electrophoretic mobility shift assays show that vertebrate GAF binds to GAGA sites in vitro. Finally, in vivo studies by chromatin immunoprecipitation confirmed that vertebrate GAF binds to GAGA-rich DNA sequences present in hox clusters. Identification of vertebrate GAF and the presence of its target sites at various developmentally regulated loci, including hox complexes, highlight the evolutionarily conserved components involved in developmental mechanisms across the evolutionary lineage and answer a long-standing question of the presence of vertebrate GAF. © 2010 Elsevier Inc.
Hendre P.S.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research |
Bhat P.R.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research |
Krishnakumar V.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research |
Aggarwal R.K.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research |
Donini P.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research
Genome | Year: 2011
Biotic or abiotic stress can cause considerable damage to crop plants that can be managed by building disease resistance in the cultivated gene pool through breeding for disease resistance genes (R-genes). R-genes, conferring resistance to diverse pathogens or pests share a high level of similarity at the DNA and protein levels in different plant species. This property of R-genes has been successfully employed to isolate putative resistance gene analogues (RGAs) using a PCR-based approach from new plant sources. Using a similar approach, in the present study, we have successfully amplified putative RGAs having nucleotide-binding-site leucine-rich repeats (NBS-LRR-type RGAs) from seven different sources: two cultivated coffee species (Coffea arabica L. and Coffea canephora Pierre ex. A. Froehner), four related taxa endemic to India (wild tree coffee species: Psilanthus bengalensis (Roem. & Schuttles) J.-F. Leroy, Psilanthus khasiana, Psilanthus travencorensis (Wight & Arn.) J.-F. Leroy, Psilanthus weightiana (Wall. ex Wight & Arn.) J.-F. Leroy), and a cDNA pool originally prepared from light-and drought-stressed Coffea arabica L. leaves. The total PCR amplicons obtained using NBS-LRR-specific primers from each source were cloned and transformed to construct seven independent libraries, from which 434 randomly picked clones were sequenced. In silico analysis of the sequenced clones revealed 27 sequences that contained characteristic RGA motifs, of which 24 had complete uninterrupted open reading frames. Comparisons of these with published RGAs showed several of these to be novel RGA sequences. Interestingly, most of such novel RGAs belonged to the related wild Psilanthus species. The data thus suggest the potential of the secondary gene pool as possible untapped donors of resistance genes to the present day cultivated species of coffee. © 2011 Published by NRC Research Press.
Siva A.B.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research |
Panneerdoss S.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research |
Sailasree P.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research |
Singh D.K.,Center for Cellular and Molecular Biology Council of Scientific and Industrial Research |
And 2 more authors.
PLoS ONE | Year: 2014
Background/Aims: The importance of sperm capacitation for mammalian fertilization has been confirmed in the present study via sperm metabolism. Involvement of the metabolic enzymes pyruvate dehydrogenase complex (PDHc) and its E3 subunit, dihydrolipoamide dehydrogenase (DLD) in hamster in vitro fertilization (IVF) via in vitro sperm capacitation is being proposed through regulation of sperm intracellular lactate, pH and calcium. Methodology and Principal Findings: Capacitated hamster spermatozoa were allowed to fertilize hamster oocytes in vitro which were then assessed for fertilization, microscopically. PDHc/DLD was inhibited by the use of the specific DLD-inhibitor, MICA (5-methoxyindole-2-carboxylic acid). Oocytes fertilized with MICA-treated (MT) [and thus PDHc/DLD-inhibited] spermatozoa showed defective fertilization where 2nd polar body release and pronuclei formation were not observed. Defective fertilization was attributable to capacitation failure owing to high lactate and low intracellular pH and calcium in MT-spermatozoa during capacitation. Moreover, this defect could be overcome by alkalinizing spermatozoa, before fertilization. Increasing intracellular calcium in spermatozoa pre-IVF and in defectively-fertilized oocytes, post-fertilization rescued the arrest seen, suggesting the role of intracellular calcium from either of the gametes in fertilization. Parallel experiments carried out with control spermatozoa capacitated in medium with low extracellular pH or high lactate substantiated the necessity of optimal sperm intracellular lactate levels, intracellular pH and calcium during sperm capacitation, for proper fertilization. Conclusions: This study confirms the importance of pyruvate/lactate metabolism in capacitating spermatozoa for successful fertilization, besides revealing for the first time the importance of sperm PDHc/ DLD in fertilization, via the modulation of sperm intracellular lactate, pH and calcium during capacitation. In addition, the observations made in the IVF studies in hamsters suggest that capacitation failures could be a plausible cause of unsuccessful fertilization encountered during human assisted reproductive technologies, like IVF and ICSI. Our studies indicate a role of sperm capacitation in the post-penetration events during fertilization. © 2014 Siva et al.