Investigations of Structural Requirements for BRD4 Inhibitors through Ligand- and Structure-Based 3D QSAR Approaches.

The bromodomain containing protein 4 (BRD4) recognizes acetylated histone proteins and plays numerous roles in the progression of a wide range of cancers, due to which it is under intense investigation as a novel anti-cancer drug target. In the present study, we performed three-dimensional quantitative structure activity relationship (3D-QSAR) molecular modeling on a series of 60 inhibitors of BRD4 protein using ligand- and structure-based alignment and different partial charges assignment methods by employing comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) approaches. The developed models were validated using various statistical methods, including non-cross validated correlation coefficient (r²), leave-one-out (LOO) cross validated correlation coefficient (q²), bootstrapping, and Fisher's randomization test. The highly reliable and predictive CoMFA (q² = 0.569, r² = 0.979) and CoMSIA (q² = 0.500, r² = 0.982) models were obtained from a structure-based 3D-QSAR approach using Merck molecular force field (MMFF94). The best models demonstrate that electrostatic and steric fields play an important role in the biological activities of these compounds. Hence, based on the contour maps information, new compounds were designed, and their binding modes were elucidated in BRD4 protein's active site. Further, the activities and physicochemical properties of the designed molecules were also predicted using the best 3D-QSAR models. We believe that predicted models will help us to understand the structural requirements of BRD4 protein inhibitors that belong to quinolinone and quinazolinone classes for the designing of better active compounds.

POLYAR, a new computer program for prediction of poly(A) sites in human sequences.

BACKGROUND: mRNA polyadenylation is an essential step of pre-mRNA processing in eukaryotes. Accurate prediction of the pre-mRNA 3'-end cleavage/polyadenylation sites is important for defining the gene boundaries and understanding gene expression mechanisms. RESULTS: 28761 human mapped poly(A) sites have been classified into three classes containing different known forms of polyadenylation signal (PAS) or none of them (PAS-strong, PAS-weak and PAS-less, respectively) and a new computer program POLYAR for the prediction of poly(A) sites of each class was developed. In comparison with polya_svm (till date the most accurate computer program for prediction of poly(A) sites) while searching for PAS-strong poly(A) sites in human sequences, POLYAR had a significantly higher prediction sensitivity (80.8% versus 65.7%) and specificity (66.4% versus 51.7%) However, when a similar sort of search was conducted for PAS-weak and PAS-less poly(A) sites, both programs had a very low prediction accuracy, which indicates that our knowledge about factors involved in the determination of the poly(A) sites is not sufficient to identify such polyadenylation regions. CONCLUSIONS: We present a new classification of polyadenylation sites into three classes and a novel computer program POLYAR for prediction of poly(A) sites/regions of each of the class. In tests, POLYAR shows high accuracy of prediction of the PAS-strong poly(A) sites, though this program's efficiency in searching for PAS-weak and PAS-less poly(A) sites is not very high but is comparable to other available programs. These findings suggest that additional characteristics of such poly(A) sites remain to be elucidated. POLYAR program with a stand-alone version for downloading is available at http://cub.comsats.edu.pk/polyapredict.htm.

Co-application of bio-fertilizer and salicylic acid improves growth, photosynthetic pigments and stress tolerance in wheat under drought stress.

Drought stress hampers the growth and productivity of wheat crop worldwide. Thus far, different strategies have been proposed to improve drought tolerance in wheat but the combined application of plant growth-promoting rhizobacteria formulated bio-fertilizer (BF) and salicylic acid (SA) has not been thoroughly explored yet. Therefore, a pot experiment was conducted to observe the effect of SA, BF, and their combination on wheat plants under optimal and drought stress conditions. Seeds priming was done with BF (107 CFU mL-1). After 2 weeks of germination, SA (one mM) was applied as a foliar spray. Drought stress was applied by withholding water supply at three-leaf stage (30 d old plants) for the next 15 d until soil moisture dropped to 10%. Foliar application of SA increased the bacterial population of BF significantly compared to the sole application of BF under irrigated as well as drought stress conditions. Co-application of BF and foliar spray of SA induced drought tolerance in wheat plants by enhancing plant biomass, photosynthetic pigments, relative water content and osmolytes, and activities of the defense-related system. Plants treated with SA and BF together under drought stress had significantly increased leaf water status, Chl a, Chl b, and carotenoids synthesis by 238%, 125%, 167%, and 122%, respectively. Moreover, the co-application of SA and BF showed maximum SOD, POD, APX, and CAT activities by 165%, 85%, 156%, and 169% in the leaves while 153%, 86%, 116% and 200% in roots under drought stress. Similarly, the combined treatment exhibited a pronounced decrease in MDA content by 54% while increased production of proteins and proline by 145% and 149%, respectively. Our results showed that the co-application of SA and BF induced better drought tolerance as compared with the sole application of SA or BF. The results obtained herein suggest that combined application of BF and SA can be applied to the wheat crop to greatly improve drought tolerance in field conditions.

Genome-Wide Profiling of Polyadenylation Events in Maize Using High-Throughput Transcriptomic Sequences.

Polyadenylation is an essential post-transcriptional modification of eukaryotic transcripts that plays critical role in transcript stability, localization, transport, and translational efficiency. About 70% genes in plants contain alternative polyadenylation (APA) sites. Despite availability of vast amount of sequencing data, to date, a comprehensive map of the polyadenylation events in maize is not available. Here, 9.48 billion RNA-Seq reads were analyzed to characterize 95,345 Poly(A) Clusters (PAC) in 23,705 (51%) maize genes. Of these, 76% were APA genes. However, most APA genes (55%) expressed a dominant PAC rather than favoring multiple PACs equally. The lincRNA genes with PACs were significantly longer in length than the genes without any PAC and about 48% genes had APA sites. Heterogeneity was observed in 52% of the PACs supporting the imprecise nature of the polyadenylation process. Genomic distribution revealed that the majority of the PACs (78%) were located in the genic regions. Unlike previous studies, large number of PACs were observed in the intergenic (n = 21,264), 5'-UTR (735), CDS (2,542), and the intronic regions (12,841). The CDS and introns with PACs were longer in length than without PACs, whereas intergenic PACs were more often associated with transcripts that lacked annotated 3'-UTRs. Nucleotide composition around PACs demonstrated AT-richness and the common upstream motif was AAUAAA, which is consistent with other plants. According to this study, only 2,830 genes still maintained the use of AAUAAA motif. This large-scale data provides useful insights about the gene expression regulation and could be utilized as evidence to validate the annotation of transcript ends.