ABSTRACT
DNA primases are pivotal enzymes in chromosomal DNA replication in all organisms. In this article, we report unique mechanistic characteristics of recombinant DNA primase from Bacillus anthracis. The mechanism of action of B. anthracis DNA primase (DnaG(BA)) may be described in several distinct steps as follows. Its mechanism of action is initiated when it binds to single-stranded DNA (ssDNA) in the form of a trimer. Although DnaG(BA) binds to different DNA sequences with moderate affinity (as expected of a mobile DNA binding protein), we found that DnaG(BA) bound to the origin of bacteriophage G4 (G4ori) with approximately 8-fold higher affinity. DnaG(BA) was strongly stimulated (>or=75-fold) by its cognate helicase, DnaB(BA), during RNA primer synthesis. With the G4ori ssDNA template, DnaG(BA) formed short (Subject(s)
Bacillus anthracis/enzymology
, DNA Primase/chemistry
, DNA Primase/metabolism
, DNA-Binding Proteins/chemistry
, Amino Acid Sequence
, Bacillus anthracis/genetics
, DNA Primase/genetics
, DNA Primers/biosynthesis
, DNA Primers/chemistry
, DNA Primers/genetics
, DNA Replication/genetics
, DNA-Binding Proteins/genetics
, DNA-Binding Proteins/metabolism
, Escherichia coli Proteins/biosynthesis
, Escherichia coli Proteins/chemistry
, Escherichia coli Proteins/genetics
, Molecular Sequence Data
, Protein Binding/genetics
, Recombinant Proteins/chemistry
, Recombinant Proteins/metabolism
, Sequence Homology, Amino Acid
