Bypass of a primase requirement for bacteriophage T4 DNA replication in vivo by a recombination enzyme, endonuclease VII.
A primase, the product of phage T4 gene 61, is required to initiate synthesis of Okazaki pieces and to allow bidirectional replication from several T4 origins. However, primase-defective T4 gene 61 mutants are viable. In these mutants, leading-strand DNA synthesis starts at the same time as in wild type infections, but, in contrast to wild type, initiation is unidirectional and the first replicative intermediates are large displacement loops. Rapid double-strand DNA replication occurs later after infection, generating multiple branched concatemers, which are cut and packaged into viable progeny particles, as in wild-type T4. Evidence is presented that this late double-strand DNA replication requires functional endonuclease VII (endo VII), the product of the T4 gene 49. We propose that endo VII can provide a backup mechanism when primase is defective, because it cuts recombinational junctions, generating 3' ends. These ends can prime DNA synthesis to copy the DNA strands that had been displaced during the initial origin-dependent replication. We explain the DNA-delay phenotype and the commonly observed temperature dependence of DNA replication in primase-deficient gene 61 mutants as a consequence of temperature-dependent translational control of gene 49 expression. In the presence or absence of functional primase endo VII is essential for correct packaging of DNA. The powerful selection that keeps the function of endo VII and expression of its gene at levels that are optimal for T4 development determines both the efficiency and the limitations of the bypass mechanism.
Mosig
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G
Department of Molecular Biology, Vanderbilt University, Nashville, TN 37235.
Luder
A
A
Ernst
A
A
Canan
N
N
D001483:Base Sequence; D019915:DNA Primase; D004261:DNA Replication; D004279:DNA, Viral; D004706:Endodeoxyribonucleases / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010957:Plasmids; D012316:RNA Nucleotidyltransferases / Q000502:physiology*; D011995:Recombination, Genetic; D013604:T-Phages / Q000235:genetics*; D014779:Virus Replication / Q000502:physiology*
D004279:DNA, Viral; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D004706:Endodeoxyribonucleases; C035571:endodeoxyribonuclease VII