| Autor: |
Bagale SS; Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India., Deshmukh PU; Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India., Lad SB; Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India., Sudarsan A; Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India., Sudhakar S; Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India., Mandal S; Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India., Kondabagil K; Department of Biosciences and Bioengineering, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India., Pradeepkumar PI; Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India. |
| Abstrakt: |
Safrole is a natural product present in many plants and plant products, including spices and essential oils. During cellular metabolism, it converts to a highly reactive trans-isosafrole (SF) intermediate that reacts with genomic DNA and forms N 2 -SF-dG and N 6 -SF-dA DNA adducts, which are detected in the oral tissue of cancer patients with betel quid chewing history. To study the SF-induced carcinogenesis and to probe the role of low fidelity translesion synthesis (TLS) polymerases in bypassing SF adducts, herein, we report the synthesis of N 2 -SF-dG modified DNAs using phosphoramidite chemistry. The N 2 -SF-dG modification in the duplex DNA does not affect the thermal stability and retains the B-form of helical conformation, indicating that this adduct may escape the radar of common DNA repair mechanisms. Primer extension studies showed that the N 2 -SF-dG adduct is bypassed by human TLS polymerases hpolκ and hpolη, which perform error-free replication across this adduct. Furthermore, molecular modeling and dynamics studies revealed that the adduct reorients to pair with the incoming nucleotide, thus allowing the effective bypass. Overall, the results indicate that hpolκ and hpolη do not distinguish the N 2 -SF-dG adduct, suggesting that they may not be involved in the safrole-induced carcinogenicity. |
