Cellular location of Mu DNA replicas

Pato, Martin L.; Waggoner, Barbara T.

doi:10.1128/jvi.38.1.249-255.1981

Cited by 16 publications

(5 citation statements)

References 27 publications

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“…Replication of full‐length (37 kb) Mu DNA in induced lysogens proceeds semi‐discontinuously (Higgins et al ., 1983), with DNA synthesis in vivo initiating 80–90% of the time at the left end of full‐length Mu (Wijffelman and van de Putte, 1977; Goosen, 1978; Pato and Waggoner, 1987). However, initiation of mini‐Mu replication in vivo takes place at the left end only ∼50% of the time (Harshey et al ., 1982; Résibois et al ., 1982a,b, 1984).…”

Section: Resultsmentioning

confidence: 99%

The phi X174-type primosome promotes replisome assembly at the site of recombination in bacteriophage Mu transposition

Jones

Nakai

1997

The EMBO Journal

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Initiation of Escherichia coli DNA synthesis primed by homologous recombination is believed to require the φX174-type primosome, a mobile priming apparatus assembled without the initiator protein DnaA. We show that this primosome plays an essential role in bacteriophage Mu DNA replication by transposition. Upon promoting transfer of Mu ends to target DNA, the Mu transpososome undergoes transition to a prereplisome that permits initiation of DNA synthesis only in the presence of primosome assembly proteins PriA, DnaT, DnaB and DnaC. These assembly proteins promote the engagement of primase and DNA polymerase III holoenzyme, initiating semi-discontinuous replication preferentially at the Mu left end. The results indicate that these proteins play a crucial role in promoting replisome assembly on a recombination intermediate.

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Section: Resultsmentioning

confidence: 99%

The phi X174-type primosome promotes replisome assembly at the site of recombination in bacteriophage Mu transposition

Jones

Nakai

1997

The EMBO Journal

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“…5). The precise mechanism of phage Mu DNA replication remains unknown, although it appears that repeated integration into host DNA is an integral part of the process (24,25,34). In spite of some differences, it is generally thought that the mechanisms of both lysogenic and lytic integration are similar (16,17,35,37).…”

Section: Discussionmentioning

confidence: 99%

“…Mu DNA are very closely linked processes (17,35). Random integration into host DNA occurs both in lysogen formation (2, 39) and continually during lytic growth (24,25,34). Despite extensive searching, phage mutants which replicate but do not integrate have never been found (31).…”

Section: Replication and Integration Of Bacteriophagementioning

confidence: 99%

Involvement of Escherichia coli K-12 DNA polymerase I in the growth of bacteriophage Mu

McBeth¹,

Taylor²

1983

J Virol

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We examined several aspects of bacteriophage Mu development in Escherichia coli strains that carry mutations in the polA structural gene for DNA polymerase I (Poll). We found that polA mutants were markedly less efficient than PolI wildtype (PolI+) strains in their capacity to form stable Mu lysogens and to support normal lytic growth of phage Mu. The frequency of lysogenization was determined for polA mutants and their isogenic PolI+ derivatives, with the result that mutants were lysogenized 3 to 8 times less frequently than were PolI+ cells. In one-step growth experiments, we found that phage Mu grew less efficiently in polA cells than in PolI+ cells, as evidenced by a 50 to 100% increase in the latent period and a 20 to 40% decrease in mean burst size in mutant cells. A further difference noted in infected polA strains was a 10-fold reduction in the frequency of Mu-mediated transposition of chromosomal genes to an F plasmid. Pulse labeling and DNA-DNA hybridization assays to measure the rate of phage Mu DNA synthesis after the induction of thermosensitive prophages indicated that phage Mu replication began at about the same time in both polA and PolI+ strains, but proceeded at a slower rate in polA cells. We conclude that Poll is normally involved in the replication and integration of phage Mu. However, since phage Mu does not exhibit an absolute requirement for normal levels of Poll, it appears that residual Poll activity in the mutant strains, other cellular enzymes, or both can partially compensate for the absence of normal Poll activity. Source' (reference) C. C. Richardson, strain D110 (30) This work This work This work This work This work This work This work CGSC, strain 5924 (23, 26) CGSC, strain 4916 (8) CGSC, strain 5642 (20) CGSC, strain 5644 (20) CGSC, strain

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“…Hybridization assays were done to determine the proportions of phage Mu and E. coli DNA in the total DNA synthesized during infection of strains AT3706 (dnaA46) and AT3822 (dnaA+) by Mucts62. The technique used was a modification of the Denhardt (2) procedure described by Kuempel (9) and used extensively for analysis of phage Mu DNA replication (19,28,29).…”

Section: E177mentioning

confidence: 99%

Growth of bacteriophage Mu in Escherichia coli dnaA mutants

McBeth¹,

Taylor²

1982

J Virol

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In one-step growth experiments we found that bacteriophage Mu grew less efficiently in nonreplicating dnaA mutants than in dnaA+ strains of Escherichia coli. Phage development in dnaA hosts was characterized by latent periods that were 15 to 30 min longer and an average burst size that was reduced by 1.5- to 4-fold. The differences in phage Mu development in dnaA and dnaA+ strains were most pronounced in cells infected at a low multiplicity and became less pronounced in cells infected at a high multiplicity. Many of these differences could be eliminated by allowing the arrested dnaA cells to restart chromosome replication just before infection. In continuous labeling experiments we found that infected dnaA strains incorporated 5 to 40 times more [methyl-3H]thymidine than did uninfected cells, depending on the multiplicity of infection. DNA-DNA hybridization assays showed that greater than 90% of this label was contained in phage Mu DNA sequences and that only small amounts of the label appeared in E. coli sequences. In contrast, substantial amounts of label were incorporated into both host and viral DNA sequences in infected dnaA+ cells. Although our results indicated that phage Mu development is not absolutely dependent on concurrent host chromosomal DNA replication, they did strongly suggest that host replication is necessary for optimal growth of this phage.

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Cellular location of Mu DNA replicas

Cited by 16 publications

References 27 publications

The phi X174-type primosome promotes replisome assembly at the site of recombination in bacteriophage Mu transposition

The phi X174-type primosome promotes replisome assembly at the site of recombination in bacteriophage Mu transposition

Involvement of Escherichia coli K-12 DNA polymerase I in the growth of bacteriophage Mu

Growth of bacteriophage Mu in Escherichia coli dnaA mutants

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