Modular architecture of the bacteriophage T7 primase couples RNA primer synthesis to DNA synthesis

Masato Kato, Takuhiro Ito, Gerhard Wagner, Charles C. Richardson, Tom Ellenberger

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

DNA primases are template-dependent RNA polymerases that synthesize oligoribonucleotide primers that can be extended by DNA polymerase. The bacterial primases consist of zinc binding and RNA polymerase domains that polymerize ribonucleotides at templating sequences of single-stranded DNA. We report a crystal structure of bacteriophage T7 primase that reveals its two domains and the presence of two Mg2+ ions bound to the active site. NMR and biochemical data show that the two domains remain separated until the primase binds to DNA and nucleotide. The zinc binding domain alone can stimulate primer extension by T7 DNA polymerase. These findings suggest that the zinc binding domain couples primer synthesis with primer utilization by securing the DNA template in the primase active site and then delivering the primed DNA template to DNA polymerase. The modular architecture of the primase and a similar mechanism of priming DNA synthesis are likely to apply broadly to prokaryotic primases.

Original languageEnglish (US)
Pages (from-to)1349-1360
Number of pages12
JournalMolecular Cell
Volume11
Issue number5
DOIs
StatePublished - May 1 2003

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DNA Primase
Bacteriophage T7
DNA
DNA-Directed DNA Polymerase
Zinc
DNA-Directed RNA Polymerases
Catalytic Domain
Oligoribonucleotides
Ribonucleotides
Single-Stranded DNA
RNA primers
Nucleotides
Ions

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Modular architecture of the bacteriophage T7 primase couples RNA primer synthesis to DNA synthesis. / Kato, Masato; Ito, Takuhiro; Wagner, Gerhard; Richardson, Charles C.; Ellenberger, Tom.

In: Molecular Cell, Vol. 11, No. 5, 01.05.2003, p. 1349-1360.

Research output: Contribution to journalArticle

Kato, Masato ; Ito, Takuhiro ; Wagner, Gerhard ; Richardson, Charles C. ; Ellenberger, Tom. / Modular architecture of the bacteriophage T7 primase couples RNA primer synthesis to DNA synthesis. In: Molecular Cell. 2003 ; Vol. 11, No. 5. pp. 1349-1360.
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