title: Streptococcus pneumoniae DNA polymerase I lacks 3'-to-5' exonuclease activity: localization of the 5'-to-3' exonucleolytic domain.
abstract: The Streptococcus pneumoniae polA gene was altered at various positions by deletions and insertions. The polypeptides encoded by these mutant polA genes were identified in S. pneumoniae. Three of them were enzymatically active. One was a fused protein containing the first 11 amino acid residues of gene 10 from coliphage T7 and the carboxyl-terminal two-thirds of pneumococcal DNA polymerase I; it possessed only polymerase activity. The other two enzymatically active proteins, which contained 620 and 351 amino acid residues from the amino terminus, respectively, lacked polymerase activity and showed only exonuclease activity. These two polymerase-deficient proteins and the wild-type protein were hyperproduced in Escherichia coli and purified. In contrast to the DNA polymerase I of Escherichia coli but similar to the corresponding enzyme of Thermus aquaticus, the pneumococcal enzyme appeared to lack 3'-to-5' exonuclease activity. The 5'-to-3' exonuclease domain was located in the amino-terminal region of the wild-type pneumococcal protein. This exonuclease activity excised deoxyribonucleoside 5'-monophosphate from both double- and single-stranded DNAs. It degraded oligonucleotide substrates to a decameric final product.
authors.affiliation: Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Velázquez, Madrid, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Diaz
authors.forename: M E
authors.initials: ME
authors.lastname: Pons
authors.forename: S A
authors.initials: SA
authors.lastname: Lacks
authors.forename: P
authors.initials: P
authors.lastname: Lopez
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D004256:DNA Polymerase I / Q000737:chemistry / Q000378:metabolism*; D004261:DNA Replication; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D016415:Sequence Alignment; D013296:Streptococcus pneumoniae / Q000201:enzymology*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D004256:DNA Polymerase I