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Full-Text Articles in Chemistry
Dna Polymerase Iii Holoenzyme From Thermus Thermophilus Identification Expression Purification Of Components And Use To Reconstitute A Processive Replicase, James M. Bullard, Jennifer C. Williams, Wendy K. Acker, Carsten Jacobi, Nebojsa Janjic, Charles S. Mchenry
Dna Polymerase Iii Holoenzyme From Thermus Thermophilus Identification Expression Purification Of Components And Use To Reconstitute A Processive Replicase, James M. Bullard, Jennifer C. Williams, Wendy K. Acker, Carsten Jacobi, Nebojsa Janjic, Charles S. Mchenry
Chemistry Faculty Publications and Presentations
DNA replication in bacteria is performed by a specialized multicomponent replicase, the DNA polymerase III holoenzyme, that consist of three essential components: a polymerase, the β sliding clamp processivity factor, and the DnaX complex clamp-loader. We report here the assembly of the minimal functional holoenzyme from Thermus thermophilus (Tth), an extreme thermophile. The minimal holoenzyme consists of α (pol III catalytic subunit), β (sliding clamp processivity factor), and the essential DnaX (τ/γ), δ and δ′ components of the DnaX complex. We show with purified recombinant proteins that these five components are required for rapid and processive DNA synthesis …
A Three-Domain Structure For The Delta Subunit Of The Dna Polymerase Iii Holoenzyme Delta Domain Iii Binds Delta' And Assembles Into The Dnax Complex, James M. Bullard, Arthur E. Pritchard, Min Sun Song, Bradley P. Glover, Anna Wieczorek, Joe Chen, Nebojsa Janjic, Charles S. Mchenry
A Three-Domain Structure For The Delta Subunit Of The Dna Polymerase Iii Holoenzyme Delta Domain Iii Binds Delta' And Assembles Into The Dnax Complex, James M. Bullard, Arthur E. Pritchard, Min Sun Song, Bradley P. Glover, Anna Wieczorek, Joe Chen, Nebojsa Janjic, Charles S. Mchenry
Chemistry Faculty Publications and Presentations
Using ψ-BLAST, we have developed a method for identifying the poorly conserved δ subunit of the DNA polymerase III holoenzyme from all sequenced bacteria. This approach, starting withEscherichia coli δ, leads not only to the identification of δ but also to the DnaX and δ′ subunits of the DnaX complex and other AAA+-class ATPases. This suggests that, although not an ATPase, δ is related structurally to the other subunits of the DnaX complex that loads the β sliding clamp processivity factor onto DNA. To test this prediction, we aligned δ sequences with those of δ′ and, using …