Origin licensing and firing require the coordinated action of multiple protein complexes. Origin licensing and firing require the coordinated action of multiple protein complexes. This cartoon depicts the steps and essential proteins involved in DNA replication origin licensing and firing, drawing from knowledge gained by studying yeast and human systems. Origin licensing occurs in the late M and G1 phase of the cell cycle when ORC1-6 binds to origins of replication. Together, CDC6 and ORC recruit CDT1 and MCM2-7, loading two MCM2-7 complexes onto dsDNA. In budding yeast, CDT1 binds to the MCM2-7 hexamer; however, these proteins have not been identified in a soluble complex without DNA in human cells. As cells transition into the S phase, Treslin, CDC45 and MTBP are recruited in a DDK-dependent manner. Pol ε and the GINS complex are recruited together with TopBP1 and RECQL4 in a CDK-dependent manner. During the S phase, licensing of additional origins is prevented by multiple mechanisms including sequestering of CDT1 by GMNN. Activation of DNA replication or origin firing requires MCM10, which aids in the bypass of the two CMG helicases past each other. MCM10 and AND-1 anchor pol α-primase to initiate DNA synthesis. As the CMG helicases progress in opposite directions, two replication forks form with pol α-primase, pol δ, pol ε and PCNA to promote DNA synthesis. Pol ε synthesizes the leading strand, while RPA binds single-stranded DNA on the lagging strand template until Okazaki fragments are produced by the consecutive action of pol α-primase and pol δ. Abbreviations: ORC, origin recognition complex; CDC, cell division cycle; CDT1, chromatin licensing and DNA replication factor 1; MCM, minichromosome maintenance; MTBP, Mdm2-binding protein; DDK, Dbf4-dependent kinase; GINS, go-ichi-ni-san; TopBP1, DNA topoisomerase II binding protein 1; RECQL4, ATP-dependent DNA helicase Q4; CDK, cyclin-dependent kinases; GMNN, geminin; AND-1, acidic nucleoplasmic DNA-binding protein 1; PCNA, proliferating cell nuclear antigen; RPA, replication protein A.