Identification and characterization of the human ORC6 homolog

Abstract

A new protein was cloned and identified as the sixth member of the human Origin Recognition Complex (ORC). The newly identified 30-kDa protein hsORC6 is 28% identical and 49% similar to ORC6p from Drosophila melanogaster, which is consistent with the identities and similarities found among the other ORC members reported in the two species. The human ORC6 gene is located on chromosome 16q12. ORC6 protein level did not change through the cell cycle. Like ORC1, ORC6 did not co-immunoprecipitate with other ORC subunits but was localized in the nucleus along with the other ORC subunits. Several cellular proteins co-immunoprecipitated with ORC6, including a 65-kDa protein that was hyperphosphorylated in G₁ and dephosphorylated in mitosis. Therefore, unlike the tight stoichiometric association of six yeast ORC subunits in one holo-complex, only a small fraction of human ORC1 and ORC6 is likely to be associated with a subcomplex of ORC2, 3, 4 and 5 suggesting differences in the architecture and regulation of human ORC.