Functional characterization of the genes coding for the terminal protein and DNA polymerase from bacteriophage GA-1. Evidence for a sliding-back mechanism during protein-primed GA-1 DNA replication

Journal of Molecular Biology
B IllanaM Salas

Abstract

We have determined the nucleotide sequence of 2676 bp at the left part of the linear genome of Bacillus subtilis bacteriophage GA-1. Computer analysis revealed that this fragment contains two open reading frames (ORFs), ORF1 and ORF2, which contain 265 and 578 codons, respectively. Data base search revealed that ORF1 and ORF2 might encode proteins similar to the terminal protein (TP) and DNA polymerase, respectively, of bacteriophage phi29. By using extracts of B. subtilis infected with GA-1, we demonstrated that GA-1 DNA replication occurs by a protein-priming mechanism in which these two viral proteins are involved. Butylphenyl dGTP (BuPdGTP), a specific inhibitor of eukaryotic-type (family B) DNA polymerases, inhibited both the protein-primed initiation step and DNA polymerization during GA-1 DNA replication. These results suggest the involvement of a eukaryotic-type DNA polymerase, probably the product of the viral ORF2, in both stages of a replication process in which the TP primes replication at both DNA ends (replication origins). Using synthetic oligonucleotides, we carried out a mutational analysis of the GA-1 DNA right end to determine the initiation site for replication. The results indicate that initiation of replic...Continue Reading

Citations

Nov 18, 2008·Proceedings of the National Academy of Sciences of the United States of America·Elisa LongásMargarita Salas
Oct 24, 2012·Proceedings of the National Academy of Sciences of the United States of America·Modesto Redrejo-RodríguezMargarita Salas
Apr 25, 2000·Nucleic Acids Research·I GascónM Salas
Aug 3, 2010·Journal of Bacteriology·Stephan KolkenbrockSusanne Fetzner
Nov 8, 2001·Journal of Bacteriology·J A HorcajadasM Salas
May 31, 2001·Microbiology and Molecular Biology Reviews : MMBR·W J MeijerM Salas
Aug 23, 2016·Frontiers in Molecular Biosciences·Margarita SalasMiguel de Vega
Apr 4, 2017·Nucleic Acids Research·Chien-Chin YangCarton W Chen
Sep 25, 2015·The Journal of Biological Chemistry·Alicia Del PradoMargarita Salas
Mar 14, 2018·Proceedings of the National Academy of Sciences of the United States of America·Alicia Del PradoMiguel de Vega
Aug 26, 2003·The Journal of Biological Chemistry·Aniko V PaulEckard Wimmer
Sep 28, 2000·The Journal of Biological Chemistry·V Gonzalez-HuiciJ M Hermoso
Oct 10, 2013·Molecular Microbiology·Modesto Redrejo-RodríguezMargarita Salas
Aug 12, 2003·Journal of Molecular Biology·Emmanuelle DufourMargarita Salas

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