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J. Biol. Chem., Vol. 263, Issue 16, 7478-7486, 06, 1988
EK Spicer, J Rush, C Fung, LJ Reha-Krantz, JD Karam and WH Konigsberg
Bacteriophage T4 gene 43 codes for the viral DNA polymerase. We report here
the sequence of gene 43 and about 70 nucleotides of 5'- and 3'- flanking
sequences, determined by both DNA and RNA sequencing. We have also purified
T4 DNA polymerase from T4 infected Escherichia coli and from E. coli
containing a gene 43 overexpression vector. A major portion of the deduced
amino acid sequence has been verified by peptide mapping and sequencing of
the purified DNA polymerase. All these results are consistent with T4 DNA
polymerase having 898 amino acids with a calculated Mr = 103,572.
Comparison of the primary structure of T4 DNA polymerase with the sequence
of other procaryotic and eucaryotic DNA polymerases indicates that T4 DNA
polymerase has regions of striking similarity with animal virus DNA
polymerases and human DNA polymerase alpha. Surprisingly, T4 DNA polymerase
shares only limited similarity with E. coli polymerase I and no detectable
similarity with T7 DNA polymerase. Based on the location of specific
mutations in T4 DNA polymerase and the conservation of particular sequences
in T4 and eucaryotic DNA polymerases, we propose that the NH2-terminal half
of T4 DNA polymerase forms a domain that carries out the 3'----5'
exonuclease activity whereas the COOH-terminal half of the polypeptide
contains the dNTP-binding site and is necessary for DNA synthesis.
Primary structure of T4 DNA polymerase. Evolutionary relatedness to eucaryotic and other procaryotic DNA polymerases
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06510-8024.
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