Extremophiles

, Volume 17, Issue 3, pp 453–461

Thermococcus kodakarensis has two functional PCNA homologs but only one is required for viability

  • Miao Pan
  • Thomas J. Santangelo
  • Ľubomíra Čuboňová
  • Zhuo Li
  • Harlette Metangmo
  • Jane Ladner
  • Jerard Hurwitz
  • John N. Reeve
  • Zvi Kelman
Original Paper

DOI: 10.1007/s00792-013-0526-8

Cite this article as:
Pan, M., Santangelo, T.J., Čuboňová, Ľ. et al. Extremophiles (2013) 17: 453. doi:10.1007/s00792-013-0526-8

Abstract

Proliferating cell nuclear antigen (PCNA) monomers assemble to form a ring-shaped clamp complex that encircles duplex DNA. PCNA binding to other proteins tethers them to the DNA providing contacts and interactions for many other enzymes essential for DNA metabolic processes. Most eukarya and euryarchaea have only one PCNA homolog but Thermococcus kodakarensis uniquely has two, designated PCNA1 and PCNA2, encoded by TK0535 and TK0582, respectively. Here, we establish that both PCNA1 and PCNA2 form homotrimers that stimulate DNA synthesis by archaeal DNA polymerases B and D and ATP hydrolysis by the replication factor C complex. In exponentially growing cells, PCNA1 is abundant and present at an ~100-fold higher concentration than PCNA2 monomers. Deletion of TK0582 (PCNA2) had no detectable effects on viability or growth whereas repeated attempts to construct a T. kodakarensis strain with TK0535 (PCNA1) deleted were unsuccessful. The implications of these observations for PCNA1 function and the origin of the two PCNA-encoding genes in T. kodakarensis are discussed.

Keywords

Archaea DNA replication Genetics PCNA Structure Thermococcus kodakarensis 

Abbreviations

PolB

DNA polymerase B

PolD

DNA polymerase D

PCNA

Proliferating cell nuclear antigen

RFC

Replication factor C

Supplementary material

792_2013_526_MOESM1_ESM.pdf (470 kb)
Supplementary material 1 (PDF 469 kb)
792_2013_526_MOESM2_ESM.pdf (67 kb)
Supplementary material 2 (PDF 66 kb)

Copyright information

© Springer Japan (outside the USA)  2013

Authors and Affiliations

  • Miao Pan
    • 1
  • Thomas J. Santangelo
    • 2
  • Ľubomíra Čuboňová
    • 2
  • Zhuo Li
    • 1
  • Harlette Metangmo
    • 1
  • Jane Ladner
    • 1
    • 3
  • Jerard Hurwitz
    • 4
  • John N. Reeve
    • 2
  • Zvi Kelman
    • 1
    • 3
  1. 1.Institute for Bioscience and Biotechnology ResearchRockvilleUSA
  2. 2.Department of MicrobiologyOhio State UniversityColumbusUSA
  3. 3.National Institute of Standards and TechnologyRockvilleUSA
  4. 4.Program of Molecular BiologyMemorial Sloan Kettering Cancer CenterNew YorkUSA

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