Biologia Molecular: Acesso Aberto

Molecular Behavior of CENP-A and PCNA throughout the Cell Cycle in Living Human HT-1080 Cells

Abstract

Kenji Sugimoto

Centromere protein A (CENP-A) is a centromere-specific histone H3 variant present in prekinetochores in nuclei and kinetochores of mitotic chromosomes. The proliferating cell nuclear antigen (PCNA) forms replication foci in S phase but diffuses in nuclei except for nucleoli in other phases. Here to elucidate the molecular behavior of CENP-A and PCNA throughout the cell cycle, we performed a prolong time-lapse imaging of HT-1080 cell line stably expressing mKO-CENP-A and EGFP-PCNA with a highly sensitive EM-CCD camera. We followed the mKO-CENP-A dots in interphase nuclei from G1 through to G2 phase, monitoring the cell cycle progression by the appearance of replication foci of EGFP-PCNA in S phase.