Ultrastructural aspects of the DNA polymerase distribution during the cell cycle

Giovanna Lattanzi, Angela Galanzi, Pietro Gobbi, Mirella Falconi, Alessandro Matteucci, Lorenzo Breschi, Marco Vitale, Giovanni Mazzotti

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

We studied the nuclear topography of the replicating enzyme DNA polymerase α in HeLa cells by transmission electron microscopy and field emission in lens scanning electron microscopy. Cells were synchronized at the G1/S-phase boundary and samples of the different phases of the cell cycle were labeled with an anti-DNA polymerase α antibody detected by an immunogold reaction. DNA synthesis was detected by immunogold labeling after bromodeoxyuridine administration. The typical labeling pattern of DNA polyroerase α observed in G1- and S-phase cells was represented by circular structures 80-100 nm in diameter surrounding an electron-dense area. In double labeled samples these circular structures were associated with bromodeoxyuridine-containing DNA replication sites, forming rosette-like structures. Field emission scanning electron microscopy performed on ultrathin cryosections revealed the chromatin fibers underlying DNA polymerase α complexes and showed that the size of the rosette-like structures corresponded to the diameter of chromatin foldings. G2- and M- phase cells showed a spread distribution of DNA polymerase α. The evidence of DNA polymerase α circular arrangement exclusively in G1- and S-phase cells, obtained by such different approaches, allowed us to consider the three-dimensional structures as DNA replication areas.

Original languageEnglish
Pages (from-to)1435-1442
Number of pages8
JournalJournal of Histochemistry and Cytochemistry
Volume46
Issue number12
Publication statusPublished - Dec 1998

Fingerprint

DNA-Directed DNA Polymerase
Cell Cycle
G1 Phase
S Phase
Bromodeoxyuridine
DNA Replication
Electron Scanning Microscopy
Chromatin
G2 Phase
Antinuclear Antibodies
DNA
Transmission Electron Microscopy
HeLa Cells
Cell Division
Lenses
Electrons
Enzymes

Keywords

  • Cell cycle
  • Chromatin structure
  • DNA polymerase α
  • DNA replication
  • Scanning electron microscopy
  • Transmission and field emission

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

Cite this

Lattanzi, G., Galanzi, A., Gobbi, P., Falconi, M., Matteucci, A., Breschi, L., ... Mazzotti, G. (1998). Ultrastructural aspects of the DNA polymerase distribution during the cell cycle. Journal of Histochemistry and Cytochemistry, 46(12), 1435-1442.

Ultrastructural aspects of the DNA polymerase distribution during the cell cycle. / Lattanzi, Giovanna; Galanzi, Angela; Gobbi, Pietro; Falconi, Mirella; Matteucci, Alessandro; Breschi, Lorenzo; Vitale, Marco; Mazzotti, Giovanni.

In: Journal of Histochemistry and Cytochemistry, Vol. 46, No. 12, 12.1998, p. 1435-1442.

Research output: Contribution to journalArticle

Lattanzi, G, Galanzi, A, Gobbi, P, Falconi, M, Matteucci, A, Breschi, L, Vitale, M & Mazzotti, G 1998, 'Ultrastructural aspects of the DNA polymerase distribution during the cell cycle', Journal of Histochemistry and Cytochemistry, vol. 46, no. 12, pp. 1435-1442.
Lattanzi G, Galanzi A, Gobbi P, Falconi M, Matteucci A, Breschi L et al. Ultrastructural aspects of the DNA polymerase distribution during the cell cycle. Journal of Histochemistry and Cytochemistry. 1998 Dec;46(12):1435-1442.
Lattanzi, Giovanna ; Galanzi, Angela ; Gobbi, Pietro ; Falconi, Mirella ; Matteucci, Alessandro ; Breschi, Lorenzo ; Vitale, Marco ; Mazzotti, Giovanni. / Ultrastructural aspects of the DNA polymerase distribution during the cell cycle. In: Journal of Histochemistry and Cytochemistry. 1998 ; Vol. 46, No. 12. pp. 1435-1442.
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