Termination within oligo(dT) tracts in template DNA by DNA polymerase gamma occurs with formation of a DNA triplex structure and is relieved by mitochondrial single-stranded DNA-binding protein.

The Journal of Biological Chemistry
V S Mikhailov, Daniel F Bogenhagen

Abstract

Xenopus laevis DNA polymerase gamma (pol gamma) exhibits low activity on a poly(dT)-oligo(dA) primer-template. We prepared a single-stranded phagemid template containing a dT41 sequence to test the ability of pol gamma to extend a primer through a defined oligo(dT) tract. pol gamma terminates in the center of this dT41 sequence. This replication arrest is abrogated by addition of single-stranded DNA-binding protein or by substitution of 7-deaza-dATP for dATP. These features are consistent with the formation of a T.A*T DNA triplex involving the primer stem. Replication arrest occurs under conditions that permit highly processive DNA synthesis by pol gamma. A similar replication arrest occurs for T7 DNA polymerase, which is also a highly processive DNA polymerase. These results suggest the possibility that DNA triplex formation can occur prior to dissociation of DNA polymerase. Primers with 3'-oligo(dA) termini annealed to a template with a longer oligo(dT) tract are not efficiently extended by pol gamma unless single-stranded DNA-binding protein is added. Thus, one of the functions of single-stranded DNA-binding protein in mtDNA maintenance may be to enable pol gamma to successfully replicate through dT-rich sequences.

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Citations

Apr 12, 2000·Medical and Veterinary Entomology·A C Lessinger, A M Azeredo-Espin
Aug 1, 1997·Antisense & Nucleic Acid Drug Development·C Giovannangeli, C Hélène
Feb 12, 2002·Journal of Virology·Victor S Mikhailov, George F Rohrmann
Mar 10, 2007·Microbiology and Molecular Biology Reviews : MMBR·Ekaterina V Mirkin, Sergei M Mirkin
Jun 11, 2004·Annual Review of Biochemistry·Laurie S Kaguni
Apr 27, 2010·Biochimica Et Biophysica Acta·Sjoerd Wanrooij, Maria Falkenberg
Mar 17, 2017·Annual Review of Biochemistry·Peter M J Burgers, Thomas A Kunkel
May 29, 2000·The Journal of Biological Chemistry·J N SpelbrinkH T Jacobs

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