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Título: | Evidence favouring the hypothesis of a conserved 3′–5′ exonuclease active site in DNA-dependent DNA polymerases |
Autor: | Blanco, Luis CSIC ORCID ; Bernad, Antonio CSIC ORCID; Salas, Margarita CSIC ORCID | Palabras clave: | Exo I Exo II and Exo III conserved segments Site-directed mutagenesis T4 DNA polymerase mutator mutants |
Fecha de publicación: | 1-mar-1992 | Editor: | Elsevier | Citación: | Gene 112(1): 139-144 (1992) | Resumen: | The complete amino acid (aa) alignment of the N-terminal domain of 33 DNA-dependent DNA polymerases encompassing the putative segments Exo I, Exo II and Exo III, proposed by Bernad et al. [Cell 59 (1989) 219–228] to form a conserved 3′–5′ exonuclease active site in prokaryotic and eukaryotic DNA polymerases, allowed us to identify and/or correct some of the most conserved segments (Exo I, II and III) in certain DNA polymerases. In particular, the aa region of T4 DNA polymerase and other eukaryotic (viral and cellular) DNA polymerases previously proposed as Exo I segment 1, did not align with the ExoI segment of Escherichia coli DNA polymerase I (PolI)-like and protein-primed DNA polymerases; instead, a new conserved region of aa similarity was identified in T4 DNA polymerase and eukaryotic (viral and cellular) DNA polymerases as their corresponding Exo I segment. Therefore according to our alignment, the recently reported T4 DNA polymerase site-directed mutants, D189A and E191A [Reha-Krantz et al., Proc. Natl. Acad. Sci. USA 88 (1991) 2417–2421], do not correspond to what we now consider the critical Exo I motif of PolI. As discussed in this communication, the functional importance of conserved segments Exo I, Exo II and Exo III is supported by site-directed mutagenesis in PolI, and in φ29, T7 and δ(Sc) DNA polymerases. Furthermore, genetically selected T4 DNA polymerase mutator mutants form two main clusters, centered in the conserved segment Exo III and in the newly identified Exo I segment. | Versión del editor: | http://dx.doi.org/10.1016/0378-1119(92)90316-H | URI: | http://hdl.handle.net/10261/39358 | DOI: | 10.1016/0378-1119(92)90316-H | ISSN: | 0378-1119 |
Aparece en las colecciones: | (CBM) Artículos |
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