Recombinant Schizosaccharomyces pombe DNA polymerase delta catalytic subunit (pol3), partial

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Cat. No.
BT32433
Source
Yeast
Synonyms
pol3; pold; SPBC336.04DNA polymerase delta catalytic subunit; EC 2.7.7.7; 3'-5' exodeoxyribonuclease; EC 3.1.11.-; DNA polymerase III
Purity
>85% (SDS-PAGE)
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Cat. No.
BT32433
Source
E.coli
Synonyms
pol3; pold; SPBC336.04DNA polymerase delta catalytic subunit; EC 2.7.7.7; 3'-5' exodeoxyribonuclease; EC 3.1.11.-; DNA polymerase III
Purity
>85% (SDS-PAGE)
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Cat. No.
BT32433
Source
E.coli
Synonyms
pol3; pold; SPBC336.04DNA polymerase delta catalytic subunit; EC 2.7.7.7; 3'-5' exodeoxyribonuclease; EC 3.1.11.-; DNA polymerase III
Purity
>85% (SDS-PAGE)
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Cat. No.
BT32433
Source
Baculovirus
Synonyms
pol3; pold; SPBC336.04DNA polymerase delta catalytic subunit; EC 2.7.7.7; 3'-5' exodeoxyribonuclease; EC 3.1.11.-; DNA polymerase III
Purity
>85% (SDS-PAGE)
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Cat. No.
BT32433
Source
Mammalian cell
Synonyms
pol3; pold; SPBC336.04DNA polymerase delta catalytic subunit; EC 2.7.7.7; 3'-5' exodeoxyribonuclease; EC 3.1.11.-; DNA polymerase III
Purity
>85% (SDS-PAGE)
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Description

Functional Insights

  • Enzymatic Activity:
    Recombinant Pol3 exhibits DNA polymerase activity only when combined with accessory proteins:

    • Proliferating Cell Nuclear Antigen (PCNA): A sliding clamp that enhances processivity .

    • Replication Factor C (RFC): Loads PCNA onto DNA in an ATP-dependent manner .
      Assays using poly(dA)/oligo(dT) templates demonstrate that partial Pol3 retains dependence on PCNA and RFC for activity, synthesizing ~20–100 nucleotides per binding event .

  • Fidelity:
    The exonuclease domain of Pol3 ensures high fidelity (~10⁻⁴ error rate), critical for maintaining genomic stability .

Production and Purification

Recombinant Pol3 is typically expressed in heterologous systems (e.g., E. coli or insect cells) and purified via:

  1. Ammonium Sulfate Precipitation: Initial enrichment of Pol3 from lysates .

  2. Chromatography: Sequential use of Q Sepharose, SP-Sepharose, and hydroxylapatite columns .

  3. Glycerol Gradient Centrifugation: Final step to isolate active fractions .

Purification StepYield (Fold)Activity (Units/mg)
Crude Extract1×0.02
Glycerol Gradient>1,000×20–25

Research Applications

  • Mechanistic Studies: Used to dissect roles in Okazaki fragment maturation and leading-strand synthesis .

  • DNA Repair Analysis: Partial Pol3 aids in understanding bypass synthesis during base excision repair (BER) and homologous recombination .

  • Evolutionary Conservation: Comparisons with human Polδ (p125 homolog) highlight conserved replication mechanisms .

Key Findings

  • Subunit Interactions: Pol3 binds Cdc1 and Cdc27 to form a stable core complex, even in recombinant systems .

  • PCNA Binding: The p22 subunit (Cdm1) dimerizes and mediates PCNA interactions, a feature conserved in human Polδ’s p12 subunit .

  • Thermosensitivity: Truncated Pol3 variants show reduced activity at elevated temperatures, aligning with cdc6 mutant phenotypes .

Open Questions

  • The functional roles of p42 and p22 in partial Pol3 constructs remain poorly characterized .

  • Structural details of the Pol3-PCNA-RFC ternary complex are unresolved.

Product Specs

Form
Lyophilized powder. We will ship the available format, but please note any format requirements when ordering, and we will accommodate your request.
Lead Time
Delivery times vary based on purchasing method and location. Consult local distributors for specific delivery times. All proteins are shipped with standard blue ice packs. Requesting dry ice will incur extra fees and requires advance notice.
Notes
Avoid repeated freeze-thaw cycles. Working aliquots can be stored at 4°C for up to one week.
Reconstitution
Briefly centrifuge the vial before opening. Reconstitute the protein in sterile deionized water to 0.1-1.0 mg/mL. Add 5-50% glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final glycerol concentration is 50%.
Shelf Life
Shelf life depends on storage conditions, buffer components, storage temperature, and protein stability. Liquid form: 6 months at -20°C/-80°C. Lyophilized form: 12 months at -20°C/-80°C.
Storage Condition
Store at -20°C/-80°C upon receiving. Aliquot for multiple uses. Avoid repeated freeze-thaw cycles.
Tag Info
The tag type is determined during manufacturing. If you have a specific tag type requirement, please inform us, and we will prioritize its development.
Synonyms
pol3; pold; SPBC336.04DNA polymerase delta catalytic subunit; EC 2.7.7.7; 3'-5' exodeoxyribonuclease; EC 3.1.11.-; DNA polymerase III
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Protein Length
Partial
Purity
>85% (SDS-PAGE)
Species
Schizosaccharomyces pombe (strain 972 / ATCC 24843) (Fission yeast)
Target Names
pol3
Uniprot No.
P30316

Target Background

Function
DNA polymerase delta's catalytic subunit (DNA polymerase III) is involved in chromosomal DNA replication, specifically lagging strand synthesis. It binds at/near replication origins and moves along DNA with the replication fork, also participating in leading strand synthesis during initiation and termination. It possesses 3'-5' proofreading exonuclease activity to correct replication errors.
Gene References Into Functions
These and previous data in budding yeast indicate that the primary functions of Pol-delta(cdc6) and Pol-epsilon(cdc20) at the eukaryotic replication fork are evolutionarily conserved. (PMID: 22144917)
Database Links

KEGG: spo:SPBC336.04

STRING: 4896.SPBC336.04.1

Protein Families
DNA polymerase type-B family
Subcellular Location
Nucleus.

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