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DNA Polymerases - The Dna Polymerase Mechanism

chain nucleotide template growing

All DNA polymerases share a common mechanism for DNA chain synthesis. The polymerization of DNA occurs by the linkage of one nucleotide at a time to the end of a preexisting DNA chain. The sequence fluctuations of the nucleotides on the DNA template upon which the DNA polymerase is moving determines which nucleotide is added onto the end of the growing DNA chain. If a thymine (T) nucleotide is positioned in the DNA template, for example, then an adenine (A) is polymerized onto the DNA chain opposite the thymine in the DNA template. If a guanine (G) nucleotide is positioned in the template, a cytosine (C) is linked to the growing DNA chain opposite the guanine. This polymerization process results in the synthesis of a DNA chain that is complementary, rather than identical, to the template strand of DNA, and is sequenced according to the proper Watson-Crick nucleotide base pairing rules. During replication, both strands of the duplex DNA molecule serve as templates. The DNA strands are separated, and each of the DNA strands is copied by the DNA polymerases. This process results in two identical copies of the original duplex DNA molecule being produced for the two cells.

The DNA polymerase uses the nucleoside triphosphate form of the deoxynucleotides to build the DNA polymer. The monophosphate form of the deoxynucleotide is incorporated into the growing DNA chain, and a pyrophosphate molecule, a kind of salt, is released. The DNA polymerase can add nucleotides only to the 3′-OH end of the growing DNA chain (see above diagram). DNA polymerase attaches an incoming triphosphate nucleotide to the 3′ position on the growing chain. Removal of a water molecule between the two attaches them, and the two terminal phosphates break off. Therefore, DNA polymerization occurs in only one direction. Some DNA polymerases are highly processive, polymerizing many nucleotides to the 3′ end of the DNA chain before falling off the DNA template. Other DNA polymerases are distributive in nature, incorporating just one nucleotide and then falling off the DNA template.

Occasionally, the DNA polymerase will incorrectly polymerize a nucleotide onto the growing DNA chain. Removal of this misinserted nucleotide must be performed by a "proofreading" exonuclease, which is a substance that removes nucleotides from the 3′ end of the DNA molecule. The combined actions of DNA polymerases and proofreading exonucleases improve the accuracy of DNA synthesis and thus minimize introduction of errors into the genome.

DNA Polymerases - The Variety Of Dna Polymerases [next]

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