Refining the domain architecture model of the replication origin firing factor Treslin/TICRR

Faithful genome duplication requires appropriately controlled replication origin firing. The metazoan origin firing regulation hub Treslin/TICRR and its yeast orthologue Sld3 share the Sld3-Treslin domain and the adjacent TopBP1/Dpb11 interaction domain. We report a revised domain architecture model of Treslin/TICRR. Protein sequence analyses uncovered a conserved Ku70-homologous beta-barrel fold in the Treslin/TICRR middle domain (M domain) and in Sld3. Thus, the Sld3-homologous Treslin/TICRR core comprises its three central domains, M domain, Sld3-Treslin domain, and TopBP1/Dpb11 interaction domain, flanked by nonconserved terminal domains, the CIT (conserved in Trestins) and the C terminus. The CIT includes a von Willebrand factor type A domain. Unexpectedly, MTBP, Treslin/TICRR, and Ku70/80 share the same N-terminal domain architecture, von Willebrand factor type A and Ku70-like beta-barrels, suggesting a common ancestry. Binding experiments using mutants and the Sld3-Sld7 dimer structure suggest that the Treslin/Sld3 and MTBP/Sld7 beta-barrels engage in homotypic interactions, reminiscent of Ku70-Ku80 dimerization. Cells expressing Treslin/TICRR domain mutants indicate that all Sld3-core domains and the non-conserved terminal domains fulfil important functions during origin firing in human cells. Thus, metazoa-specific and widely conserved molecular processes cooperate during metazoan origin firing.

Automated summary

The faithful duplication of the genome requires controlled replication origin firing. The proteins Treslin/TICRR and its yeast counterpart Sld3 play a crucial role in regulating the firing of replication origins. A revised domain architecture model of Treslin/TICRR is proposed, revealing a conserved beta-barrel fold in the middle domain (M domain) shared by Treslin/TICRR and Sld3. It is found that Treslin/Sld3 and MTBP/Sld7 engage in homotypic interactions similar to the dimerization of Ku70-Ku80. Mutants of the Treslin/TICRR domain indicate the importance of all Sld3-core domains and the non-conserved terminal domains during origin firing in human cells.