Interaction of Ribonucleoside Triphosphates with the Gene 4 Primase of Bacteriophage T7*

@article{Frick1999InteractionOR,
  title={Interaction of Ribonucleoside Triphosphates with the Gene 4 Primase of Bacteriophage T7*},
  author={D. Frick and Shiv Kumar and C. Richardson},
  journal={The Journal of Biological Chemistry},
  year={1999},
  volume={274},
  pages={35899 - 35907}
}
The primase fragment of bacteriophage T7 gene 4 protein catalyzes the synthesis of oligoribonucleotides in the presence of ATP, CTP, Mg2+ (or Mn2+), and DNA containing a primase recognition site. During chain initiation, ATP binds with a K m of 0.32 mm, and CTP binds with a K m of 0.85 mm. Synthesis of the dinucleotides proceeds at a rate of 3.8/s. The dinucleotide either dissociates or is extended to a tetranucleotide. The primase preferentially inserts ribonucleotides forming Watson-Crick… Expand
Interaction of Bacteriophage T7 Gene 4 Primase with Its Template Recognition Site*
TLDR
The primase fragment of the bacteriophage T7 63-kDa gene 4 helicase/primase protein contains the 271 N-terminal amino acid residues and lacks helicase activity, suggesting that the primase binds DNA in a sequence-specific manner. Expand
The heterodimeric primase of the hyperthermophilic archaeon Sulfolobus solfataricus possesses DNA and RNA primase, polymerase and 3'-terminal nucleotidyl transferase activities.
TLDR
It is revealed that the two-subunit DNA-dependent primase possesses a template-independent 3'-terminal nucleotidyl transferase activity, which is crucial for nucleic acid synthesis and residues important for the binding of free nucleotides. Expand
Mechanism of sequence-specific template binding by the DNA primase of bacteriophage T7
TLDR
A model in which conformational changes induced during primer synthesis facilitate contact between the zinc-binding domain and the polymerase domain is presented, which shows that T7 DNA primase has only a slightly higher affinity for DNA containing the primase recognition sequence than for DNA lacking the recognition site. Expand
Acidic Residues in the Nucleotide-binding Site of the Bacteriophage T7 DNA Primase*
TLDR
Despite the inability to catalyze phosphodiester bond formation, the altered proteins recognize the sequence 5′-GTC-3′ in the template and deliver preformed primer to T7 DNA polymerase. Expand
Essential Lysine Residues in the RNA Polymerase Domain of the Gene 4 Primase-Helicase of Bacteriophage T7*
TLDR
To identify the catalytic core of the T7 DNA primase, single-point mutations were introduced into a basic region that shares sequence homology with RNA polymerases. Expand
A Complex of the Bacteriophage T7 Primase-Helicase and DNA Polymerase Directs Primer Utilization*
TLDR
Results show that a single subunit of the primase-helicase hexamer contains all of the residues required for primer synthesis and for utilization of primers by T7 DNA polymerase. Expand
Structure and mechanism of human PrimPol, a DNA polymerase with primase activity
TLDR
The first structure of human PrimPol in ternary complex with a DNA template-primer and an incoming deoxynucleoside triphosphate (dNTP) is presented, addressing long-standing questions about how DNA primases actually initiate synthesis and how primase and polymerase activities combine in a single enzyme to carry out DNA synthesis. Expand
Structures of human primase reveal design of nucleotide elongation site and mode of Pol α tethering
TLDR
The crystal structure of human primase in heterodimeric form consisting of full-length catalytic subunit and a C-terminally truncated large subunit is described and it is shown that the small subunit integrates primer initiation and elongation within the same set of functional residues. Expand
Properties of an unusual DNA primase from an archaeal plasmid
TLDR
The primase activity of the replication protein from the archaeal plasmid pRN1 synthesizes a rather unusual mixed primer consisting of a single ribonucleotide at the 5′ end followed by seven deoxynucleotides. Expand
The Roles of Tryptophans in Primer Synthesis by the DNA Primase of Bacteriophage T7*
TLDR
Replacement of Trp-42, -97, or -147 with the structurally similar tyrosine disturbs the conformation of the ZBD and reduces NTP binding and the catalysis step, while replacement at position 42, 97, or 147 reduced primer synthesis, whereas substitution at position 69 or 255 did not. Expand
...
1
2
3
4
...

References

SHOWING 1-10 OF 47 REFERENCES
Interaction of Bacteriophage T7 Gene 4 Primase with Its Template Recognition Site*
TLDR
The primase fragment of the bacteriophage T7 63-kDa gene 4 helicase/primase protein contains the 271 N-terminal amino acid residues and lacks helicase activity, suggesting that the primase binds DNA in a sequence-specific manner. Expand
Gene 4 DNA Primase of Bacteriophage T7 Mediates the Annealing and Extension of Ribo-oligonucleotides at Primase Recognition Sites*
TLDR
T7 primase extends the dinucleotide AC and trinucleotide ACC to ACCC in the presence of CTP and an appropriate template, whereas other dinucleotides are extended less efficiently; the deoxyribodin nucleotide dAC is not extended. Expand
Role of bacteriophage T7 DNA primase in the initiation of DNA strand synthesis.
TLDR
With the aid of primase, T7 DNA polymerase can also utilize efficiently a variety of synthetic tri-, tetra-, or pentanucleotides as chain initiators. Expand
Template Recognition and Ribonucleotide Specificity of the DNA Primase of Bacteriophage T7*
TLDR
It is proposed that pausing is required for the incorporation of less preferred nucleotides and thatPausing is decreased by the ability of the primase to translocate 5′ to 3′ on templates having long 3′-flanking sequences. Expand
DNA-dependent nucleoside 5'-triphosphatase activity of the gene 4 protein of bacteriophage T7.
TLDR
It is concluded that the energy derived from the hydrolysis of NTPs by the gene 4 protein is required for translocation of the protein to primase recognition sites, and single-stranded DNA-dependent nucleoside 5'-triphosphatase of the gene4 protein is coupled to the unidirectional translocation. Expand
Deoxyribonucleic acid polymerase of bacteriophage T7. Characterization of the exonuclease activities of the gene 5 protein and the reconstituted polymerase.
TLDR
Hydlysis of single-stranded DNA by the gene 5 protein is inhibited even in the absence of thioredoxin where there is less than 2% active T7 DNA polymerase activity. Expand
An N-terminal fragment of the gene 4 helicase/primase of bacteriophage T7 retains primase activity in the absence of helicase activity.
TLDR
The primase fragment is defective in the interaction with T7 DNA polymerase in that primer synthesis cannot be coupled to DNA synthesis, and it catalyzes template-directed synthesis of di-, tri-, and tetranucleotides. Expand
Effects of base analog substitutions in the noncoding dC of the 3'-d(CTG)-5' template recognition site of the bacteriophage T7 primase.
TLDR
The interactions between T7 primase and the functional groups of the 3'-dC residue are examined by measuring the ability of the primase to catalyze the synthesis of oligoribonucleotides on synthetic single-stranded 20-mer templates that contain analogs of dC in the conserved pentanucleotide recognition site. Expand
Nucleotide-dependent binding of the gene 4 protein of bacteriophage T7 to single-stranded DNA.
The gene 4 protein of bacteriophage T7 is a multifunctional enzyme that catalyzes (i) the hydrolysis of nucleoside 5'-triphosphates, (ii) the synthesis of tetraribonucleotide primers at specificExpand
A Domain of the Gene 4 Helicase/Primase of Bacteriophage T7 Required for the Formation of an Active Hexamer (*)
TLDR
The data demonstrate that this region of the T7 gene 4 protein is important for the protein-protein contacts necessary for both hexamer formation and the interactions between the subunits of the hexamer required for coordinated nucleotide hydrolysis, translocation on single-stranded DNA, and unwinding of double-Stranded DNA. Expand
...
1
2
3
4
5
...