The human autoantigen MCP1 is required during early stages of DNA replication

Metaphase chromosome protein 1 (MCP1) is a nuclear autoantigen that is asso ciated with condensed chromosomes throughout mitosis. During interphase, th is antigen shows a speckle distribution in the nucleus, excluding the nucle olus. Additionally, MCP1 binds tightly to the scaffold/matrix component of nuclei and isolated chromosomes. In order to determine the in-vivo localiza tion of the antigen, we have expressed MCP1 fused to EGFP in tissue culture cells. The results demonstrate that MCP1 is located in the nucleus during interphase and during mitosis associates tightly to condensed chromosomes. Furthermore, microinjection of specific antibody confirms these results. We have used a specific monoclonal antibody (mAb 402) against MCP1 to assess the function of this antigen during cell cycle progression. HeLa and Ptk-2 cells that were microinjected into the nucleus and/or cytoplasm at G1/S and very early S phase were not able to progress and complete DNA replication. However, injection of mAb 402 at mid or late S phase does not prevent comp letion of DNA replication and subsequent progression into mitosis. Microinj ection of mAb 402 in Ptk-2 cells synchronized in mitosis did not interfere with progression of mitosis and cells divided. Our results suggest that MCP 1 is required at the G1/S transition and during early S phase.