Mapping platinum adducts on yeast ribosomal RNA using high-throughput sequencing

Area of Science:

• Biochemistry
• Molecular Biology
• Genomics

Background:

• Mapping small-molecule binding sites on cellular RNAs is crucial for understanding molecular interactions.
• Platinum(II) reagents are established DNA and RNA crosslinkers, but identifying their specific targets genome-wide within cells is difficult.

Purpose of the Study:

• To develop and apply a high-throughput sequencing method for identifying platinum-RNA adducts in vivo.
• To investigate the targets of cisplatin, a platinum(II) reagent, on ribosomal RNA (rRNA) in Saccharomyces cerevisiae.

Main Methods:

• Application of high-throughput sequencing, termed 'Pt-Seq', to analyze RNA modifications.
• Treatment of Saccharomyces cerevisiae (yeast) cells with cisplatin.
• Identification of platinum-ribosomal RNA (rRNA) adducts.

Main Results:

• Successfully mapped platinum-rRNA adducts in yeast cells.
• Demonstrated the utility of Pt-Seq for genome-wide identification of Pt targets on RNA.
• Provided insights into the interaction of cisplatin with rRNA.

Conclusions:

• High-throughput Pt-Seq is an effective method for identifying platinum-RNA adducts in cells.
• The study identified specific rRNA targets of cisplatin in S. cerevisiae.
• This approach advances the study of small-molecule interactions with RNA in a cellular context.