Journal
MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS
Volume 832, Issue -, Pages 41-51Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mrgentox.2018.05.022
Keywords
Aquatic pollution; Gene expression patterns; Mutation; p53; Rita rita
Funding
- Indian Council of Agricultural Research (ICAR), New Delhi (CIFRI Core Project) [FREM/ER/12/03/05]
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The present study was undertaken to investigate the alterations in gene expression patterns and for mutation analysis of p53 in the riverine catfish Rita rita collected from polluted riverine habitat. The partial p53 gene sequence of Rita rita generated showed a high degree of similarities with the DNA binding domains of fishes, mice and human. Transcriptomic analysis, carried out by quantitative real-time Polymerase Chain Reaction (RT-qPCR), showed significant down-regulation of p53 in fishes collected from most of the polluted stretches. Similar trend in protein abundance was observed by western blot analysis. Down-regulation of p53 was more pronounced in gill than liver. Expression patterns of p53 suggest that exposure to a multitude of contaminants in the natural riverine ecosystem could suppress the expression of p53. Genomic DNA showed a low stained smear pattern upon electrophoresis, with no evidence of DNA fragmentation. For mutation analysis PCR-SSCP followed by sequence analysis was carried out, which identified eight mutations; two at codon level and six missense mutations in the DNA binding domain IV and V. Secondary structure prediction showed that these mutations could lead to impairment of protein structure. Thus, the present study indicated that aquatic pollution has impacted these lower vertebrates which are reflected by the down-regulation of tumor suppressor protein (p53) in majority of the stretches studied.
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