This study aims at understanding the roles of Spartan in translesion synthesis (TLS) and UV-induced carcinogenesis. TLS is a post-replication repair pathway, which mediates bypass of bulky DNA lesions that stall replication forks, during DNA replication. Bypass of DNA lesions is mediated by specialized low-fidelity TLS polymerase that can replicate over distortions or bulky DNA adducts efficiently, leaving behind the lesion to be repaired at a later time point. The critical step in TLS is the switch of the replicative polymeras with TLS polymerase, which is proposed to be mediated by RAD18-dependent ubiquitination of PCNA (ub-PCNA). However, the exact mechanism is not clear. We identified Spartan as a key regulator of TLS. Depletion of Spartan renders cells sensitive to UV damage, results in a decrease in PCNA mono-ubiquitination, which is accompanied by a reduction of RAD18 chromatin association and RAD18 localization to DNA damage sites. Interestingly, Spartan binds to replicative DNA polymerase under normal conditions, but preferentially associates with TLS polymerase POLH upon UV damage. Based on our observations, we proposed that Spartan is a key regulator of TLS, required to stabilize RAD18 and ub-PCNA at the sites of DNA damage and may directly regulate the switch from replicative polymerase to TLS polymerase during TLS. However, how Spartan mediates its regulatory functions and the switch between the DNA polymerases during TLS is not known and will be the focus of this study.
The specific aims for this proposal are:
Aim 1) Determine the function of Spartan in TLS upon UV damage;
Aim 2) Determine the roles of Spartan mediated TLS in UV-induced carcinogenesis and resistance to chemotherapeutic agents;
Aim 3) Determine the in vivo physiological functions of Spartan. This proposed study will not only provide useful insights into the regulatory mechanism of TLS process, but also, will explore the potential of targeting Spartan for cancer therapy.
Translesion synthesis (TLS) is a post-replication repair pathway, which allows the replication machinery to bypass DNA lesions that block the progression of the replication fork during DNA replication. TLS employs specialized low-fidelity TLS polymerases to bypass DNA lesions, thereby preventing fork collapse and genome instability. Spartan is a PCNA-interacting protein, required for cell survival in response to UV-induced DNA lesions and may have a direct role in regulating the switch from replicative polymerase to TLS polymerase at the damage site during TLS.