Journal of Molecular Medicine

, Volume 77, Issue 4, pp 386–392

The DNA-binding subunit p140 of replication factor C is upregulated in cycling cells and associates with G1 phase cell cycle regulatory proteins

  • Heiko van der Kuip
  • Birgit Carius
  • S. Jaharul Haque
  • Bryan R. G. Williams
  • Christoph Huber
  • T. Fischer
Original Article

DOI: 10.1007/s001090050365

Cite this article as:
van der Kuip, H., Carius, B., Haque, S. et al. J Mol Med (1999) 77: 386. doi:10.1007/s001090050365

Abstract

The DNA-binding subunit of replication factor C (RFCp140) plays an important role in both DNA replication and DNA repair. The mechanisms regulating activation of RFCp140 thereby controlling replication and cellular proliferation are largely unknown. We analyzed protein expression of RFCp140 during cell cycle progression and investigated the association of RFCp140 with cell cycle regulatory proteins in cell lines of various tissue origin and in primary hematopoietic cells. Western and Northern blot analyses of RFCp140 from synchronized cells showed downregulation of RFCp140 when cells enter a G0-like quiescent state and upregulation of RFCp140 in cycling cells. Translocation from the cytoplasmic compartment to the nucleus did not account for the significant increase in RFCp140 protein levels observed in cycling cells. To investigate a potential association of RFCp140 with cell cycle regulatory proteins coimmunoprecipitation assays were performed. These studies demonstrated specific binding of RFCp140 to cdk4-kinase in hematopoietic and fibroblast cell lines. Additional coimmunoprecipitation studies revealed specific association of RFCp140 with cyclin D1, p21, proliferating cell nuclear antigen, and retinoblastoma protein. These findings link DNA replication and repair factor RFCp140 to G1 phase cell cycle regulatory elements critically involved in cell cycle control.

Key words Replication factor CRFCp140Cell cycleG1 phase regulatory proteins

Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • Heiko van der Kuip
    • 1
  • Birgit Carius
    • 1
  • S. Jaharul Haque
    • 2
  • Bryan R. G. Williams
    • 2
  • Christoph Huber
    • 1
  • T. Fischer
    • 1
  1. 1.Department of Hematology, Johannes Gutenberg University School of Medicine, Langenbeckstrasse 1, D-55131 Mainz, Germany e-mail: t.fischer@3-med.klinik.uni-mainz.de, Tel.: +49-6131-176544/177281, Fax: +49-6131-176678DE
  2. 2.Department of Cancer Biology, Research Institute, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH 44195, USAUS