Thymidine 5'-O-monophosphorothioate induces HeLa cell migration by activation of the P2Y6 receptor

Area of Science:

• Biochemistry
• Molecular Biology
• Cell Biology

Background:

• Purinergic signaling via P2Y receptors regulates critical cellular functions.
• Nucleotides such as ATP, ADP, UTP, and UDP are known P2Y receptor ligands.
• The role of nucleoside 5 -O-monophosphorothioate analogs as P2Y receptor ligands was previously unclear.

Purpose of the Study:

• To investigate the activity of nucleoside 5 -O-monophosphorothioate analogs as ligands for P2Y receptors.
• To characterize the specific interaction of thymidine 5 -O-monophosphorothioate (TMPS) with P2Y receptor subtypes.
• To explore the involvement of the P2Y6 receptor in TMPS-induced cell migration.

Main Methods:

• Synthesis and testing of nucleoside 5 -O-monophosphorothioate analogs.
• Pharmacological characterization of TMPS as a P2Y6 receptor agonist.
• Assessment of TMPS-induced HeLa cell migration.
• Validation using siRNA-mediated P2Y6 receptor knockdown and a selective P2Y6R antagonist (MRS 2578).

Main Results:

• Nucleoside 5 -O-monophosphorothioate analogs function as ligands for specific P2Y receptor subtypes.
• Thymidine 5 -O-monophosphorothioate (TMPS) acts as a specific partial agonist of the P2Y6 receptor.
• TMPS induces migration in HeLa cells, an effect dependent on P2Y6 receptor activation.
• P2Y6 receptor knockdown and antagonist treatment abolished TMPS-induced cell migration.

Conclusions:

• TMPS is a novel ligand for the P2Y6 receptor, acting as a partial agonist.
• The P2Y6 receptor mediates TMPS-induced cell migration.
• The enhanced stability and affinity of TMPS for P2Y6R may underlie long-term receptor effects.