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PMID:27052734
| Citation |
Liu, B, Gu, S, Liang, N, Xiong, M, Xue, Q, Lu, S, Hu, F and Zhang, H (2016) Pseudomonas aeruginosa phage PaP1 DNA polymerase is an A-family DNA polymerase demonstrating ssDNA and dsDNA 3'-5' exonuclease activity. Virus Genes 52:538-51 |
|---|---|
| Abstract |
Most phages contain DNA polymerases, which are essential for DNA replication and propagation in infected host bacteria. However, our knowledge on phage-encoded DNA polymerases remains limited. This study investigated the function of a novel DNA polymerase of PaP1, which is the lytic phage of Pseudomonas aeruginosa. PaP1 encodes its sole DNA polymerase called Gp90 that was predicted as an A-family DNA polymerase with polymerase and 3'-5' exonuclease activities. The sequence of Gp90 is homologous but not identical to that of other A-family DNA polymerases, such as T7 DNA polymerases (Pol) and DNA Pol I. The purified Gp90 demonstrated a polymerase activity. The processivity of Gp90 in DNA replication and its efficiency in single-dNTP incorporation are similar to those of T7 Pol with processive thioredoxin (T7 Pol/trx). Gp90 can degrade ssDNA and dsDNA in 3'-5' direction at a similar rate, which is considerably lower than that of T7 Pol/trx. The optimized conditions for polymerization were a temperature of 37 °C and a buffer consisting of 40 mM Tris-HCl (pH 8.0), 30 mM MgCl2, and 200 mM NaCl. These studies on DNA polymerase encoded by PaP1 help advance our knowledge on phage-encoded DNA polymerases and elucidate PaP1 propagation in infected P. aeruginosa. |
| Links |
PubMed Online version:10.1007/s11262-016-1329-7 |
| Keywords |
Amino Acid Sequence; Bacteriophages/genetics; DNA/genetics; DNA Replication/genetics; DNA, Single-Stranded/genetics; DNA-Directed DNA Polymerase/genetics; Exonucleases/genetics; Pseudomonas aeruginosa/genetics; Sequence Alignment; Thioredoxins/genetics |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0034061: DNA polymerase activity |
ECO:0000314: |
F |
Fig 6. Authors used a primer extension assay to show extension of a 27-mer primer to 62-mer DNA fragment over 1 minute using different concentrations of GP90 yielding 98% conversion with 20 nM of GP90. |
complete | ||||
Notes
See also
References
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