Insight into the human DNA primase interaction with template-primer

Andrey G. Baranovskiy, Yinbo Zhang, Yoshiaki Suwa, Jianyou Gu, Nigar D. Babayeva, Youri I Pavlov, Tahir H Tahirov

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

DNA replication in almost all organisms depends on the activity of DNA primase, a DNA-dependent RNA polymerase that synthesizes short RNA primers of defined size for DNA polymerases. Eukaryotic and archaeal primases are heterodimers consisting of small catalytic and large accessory subunits, both of which are necessary for the activity. The mode of interaction of primase subunits with substrates during the various steps of primer synthesis that results in the counting of primer length is not clear. Here we show that the C-terminal domain of the large subunit (p58C) plays a major role in template-primer binding and also defines the elements of the DNA template and the RNA primer that interact with p58C. The specific mode of interaction with a template-primer involving the terminal 5'-triphosphate of RNA and the 3'-overhang of DNA results in a stable complex between p58C and the DNA/RNA duplex. Our results explain how p58C participates in RNA synthesis and primer length counting and also indicate that the binding site for initiating NTP is located on p58C. These findings provide notable insight into the mechanism of primase function and are applicable for DNA primases from other species.

Original languageEnglish (US)
Pages (from-to)4793-4802
Number of pages10
JournalJournal of Biological Chemistry
Volume291
Issue number9
DOIs
StatePublished - Feb 26 2016

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DNA Primase
DNA
RNA
Accessories
DNA-Directed DNA Polymerase
DNA-Directed RNA Polymerases
DNA Replication
Binding Sites
Substrates
RNA primers

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Insight into the human DNA primase interaction with template-primer. / Baranovskiy, Andrey G.; Zhang, Yinbo; Suwa, Yoshiaki; Gu, Jianyou; Babayeva, Nigar D.; Pavlov, Youri I; Tahirov, Tahir H.

In: Journal of Biological Chemistry, Vol. 291, No. 9, 26.02.2016, p. 4793-4802.

Research output: Contribution to journalArticle

Baranovskiy, Andrey G. ; Zhang, Yinbo ; Suwa, Yoshiaki ; Gu, Jianyou ; Babayeva, Nigar D. ; Pavlov, Youri I ; Tahirov, Tahir H. / Insight into the human DNA primase interaction with template-primer. In: Journal of Biological Chemistry. 2016 ; Vol. 291, No. 9. pp. 4793-4802.
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