Accurate DNA synthesis by Sulfolobus solfataricus DNA polymerase B1 at high temperature

Zhang, Likui; Lou, Huiqiang; Guo, Li; Zhan, Zhengyan; Duan, Zengqiang; Guo, Xin; Huang, Li

doi:10.1007/s00792-009-0292-9

Cited by 8 publications

(11 citation statements)

References 51 publications

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“…It has been well documented that reaction temperature and buffer conditions such as pH and divalent metal influence the fidelity of DNA polymerases (Zhang et al 2009, 2010). A standard set of reaction conditions using ThermoPol buffer was used to be consistent with previous studies on fidelity (Mattila et al 1991) but may not reflect buffer conditions in vivo.…”

Section: Resultsmentioning

confidence: 99%

Characterization of Family D DNA polymerase from Thermococcus sp. 9°N

et al. 2014

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Accurate DNA replication is essential for maintenance of every genome. All archaeal genomes except Crenarchaea, encode for a member of Family B (polB) and Family D (polD) DNA polymerases. Gene deletion studies in Thermococcus kodakaraensis and Methanococcus maripaludis show that polD is the only essential DNA polymerase in these organisms. Thus, polD may be the primary replicative DNA polymerase for both leading and lagging strand synthesis. To understand this unique archaeal enzyme, we report the biochemical characterization of a heterodimeric polD from Thermococcus. PolD contains both DNA polymerase and proofreading 3′–5′ exonuclease activities to ensure efficient and accurate genome duplication. The polD incorporation fidelity was determined for the first time. Despite containing 3′–5′ exonuclease proofreading activity, polD has a relatively high error rate (95 × 10−5) compared to polB (19 × 10−5) and at least 10-fold higher than the polB DNA polymerases from yeast (polε and polδ) or Escherichia coli DNA polIII holoenzyme. The implications of polD fidelity and biochemical properties in leading and lagging strand synthesis are discussed.Electronic supplementary materialThe online version of this article (doi:10.1007/s00792-014-0646-9) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Characterization of Family D DNA polymerase from Thermococcus sp. 9°N

et al. 2014

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“…DNA polymerases of Thermus aquaticus , Thermococcus litoralis and S. solfataricus ). Mutant frequencies obtained with those enzymes at 70–72 °C were similar or moderately reduced (< 2‐fold) compared to estimates obtained at 55 °C using M13‐based forward mutation assays .…”

Section: Discussionmentioning

confidence: 59%

Temperature effects on the fidelity of a thermostable HIV‐1 reverse transcriptase

Saura

Menéndez‐Arias

2013

The FEBS Journal

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Transcriptomics and gene expression analysis are largely dependent of the availability of efficient thermostable reverse transcriptases (RTs). However, the intrinsic fidelity of DNA synthesis catalyzed by retroviral RTs is low. Reported error rates are in the range 1.2 × 10−5–6.7 × 10−4, with oncoretroviral RTs being the most faithful enzymes. Wild‐type HIV‐1 group O (HIV‐1O) RT is a thermostable polymerase that is able to synthesize cDNA at temperatures as high as 70 °C. At 37 °C, its error rate has been estimated at 5.8 × 10−5 in M13mp2 lacZ‐based forward mutation assays. However, at higher temperatures (e.g. 50 and 55 °C), the accuracy of HIV‐1O RT is increased by approximately two‐ to five‐fold. At 55 °C, the HIV‐1O RT error rate (1.3 × 10−5) was similar to that shown by the AffinityScript (Agilent Technologies Inc., La Jolla, CA, USA) RT, a commercially available thermostable murine leukaemia virus RT. At higher temperatures, the increased accuracy of the HIV‐1 enzyme results from a lower base substitution error rate, although it shows a higher tendency to introduce frameshifts. Kinetic studies carried out with model template‐primers suggest minor differences in nucleotide discrimination, although, at higher temperatures, HIV‐1O RT showed a reduced ability to extend mispaired template‐primers.

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“…Strong processivity of DNA pols allow them to synthesize DNA around the plasmid template in SDM. Family B archaeal DNA pols have high fidelity Zhang et al 2010a) but have different processivity depending on archaeal organisms. The processivity and activity of archaeal DNA pols is enhanced by DNA replication proteins, such as replication factor C (RFC) and proliferating cell nuclear antigen (PCNA) in archaea (Xing et al 2014;Pan et al 2011).…”

Section: Site-directed Mutagenesis By Pcrmentioning

confidence: 99%

Archaeal DNA polymerases in biotechnology

Zhang

Kang

et al. 2015

Appl Microbiol Biotechnol

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DNA polymerase (pol) is a ubiquitous enzyme that synthesizes DNA strands in all living cells. In vitro, DNA pol is used for DNA manipulation, including cloning, PCR, site-directed mutagenesis, sequencing, and several other applications. Family B archaeal DNA pols have been widely used for molecular biological methods. Biochemical and structural studies reveal that each archaeal DNA pol has different characteristics with respect to fidelity, processivity and thermostability. Due to their high fidelity and strong thermostability, family B archaeal DNA pols have the extensive application on high-fidelity PCR, DNA sequencing, and site-directed mutagenesis while family Y archaeal DNA pols have the potential for error-prone PCR and random mutagenesis because of their low fidelity and strong thermostability. This information combined with mutational analysis has been used to construct novel DNA pols with altered properties that enhance their use as biotechnological reagents. In this review, we focus on the development and use of family B archaeal DNA pols.

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Accurate DNA synthesis by Sulfolobus solfataricus DNA polymerase B1 at high temperature

Cited by 8 publications

References 51 publications

Characterization of Family D DNA polymerase from Thermococcus sp. 9°N

Characterization of Family D DNA polymerase from Thermococcus sp. 9°N

Temperature effects on the fidelity of a thermostable HIV‐1 reverse transcriptase

Archaeal DNA polymerases in biotechnology

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