Dynamics of human replication protein A subunit distribution and partitioning in the cell cycle.
Human replication protein A (RPA) is a three-sub-unit protein complex involved in DNA replication, repair, and recombination. To gain insight into the dynamics of subunit assembly, we examined the subcellular distribution of RPA subunits (p70, p34, and p11) during the cell cycle. All three subunits colocalized in G1 and S phases, showing a diffuse nuclear distribution in G1 but a dot-like nuclear pattern in S phase. During S phase, the subunits showed a pattern reminiscent of the replication granules/factories described by others as sites of replication machinery. In meta-phase, p70 preferentially associated with the spindle poles, p34 was found on chromosomes, and p11 remained in the cytoplasm. In telophase, p70 and p34 appeared in the forming daughter nuclei; p11 remained in the cytoplasm until G1. Among the three subunits only p34 was associated with the nuclear matrix and this association persisted throughout the cell cycle. We conclude that (i) RPA complex assembly is differentially regulated, (ii) the replication machinery may be anchored to the nuclear matrix, and (iii) RPA subunits partition during mitosis and sort into daughter nuclei by different routes.
Murti
K G
KG
Department of Virology and Molecular Biology St. Jude Children's Research Hospital, Memphis, Tennessee 38105-2794, USA.
He
D C
DC
Brinkley
B R
BR
Scott
R
R
Lee
S H
SH
D002453:Cell Cycle; D002467:Cell Nucleus / Q000737:chemistry*; D003593:Cytoplasm / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D007399:Interphase; D008938:Mitosis; D015530:Nuclear Matrix / Q000737:chemistry; D051716:Replication Protein A
D004268:DNA-Binding Proteins; C497167:RPA1 protein, human; D051716:Replication Protein A