Transcription of bacteriophage φX174 in vitro: Analysis with restriction enzymes

Axelrod, Nancy

doi:10.1016/s0022-2836(76)80116-7

Cited by 21 publications

(19 citation statements)

References 11 publications

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“…Lines a through e indicate the deduced sites for transcription termination obtained from in vivo and in vitro experiments and from theoretical considerations. Line a, in vivo termination sites, from Hayashi et al (10); line b, in vitro termination sites from Axelrod (2,3); line c, termination sites from Smith and Sinsheimer (30-32); line d, termination sites from Kapitza et al (18,19) (p indicates rho-dependent termination); line e, termination sites from McMahon and Tinoco (21) (numbers represent the positions on the new map of Sanger et al [27]). More detailed explanations of Fig.…”

Section: Methodsmentioning

confidence: 99%

Bacteriophage phi X174-specific mRNA synthesis in cells deficient in termination factor rho activity

Imai¹

1981

J Virol

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A previous report (Hayashi et al., Proc. Natl. Acad. Sci. U.S.A. 73:3519-3523, 1976) indicated that in vivo bacteriophage 4)X174 mRNA's terminate after genes J, F, G, and H. However, termination at these sites is not stringent. To determine whether termination of 4X174 transcription depends on rho factor activity, we introduced a temperature-sensitive rho mutation (nitA) into a 4X174-sensitive host cell line and determined termination sites in wild-type and nitA cells. We found that (i) normal OX174 terminators were recognized in 4X174-infected nitA cells, (ii) the rho mutation relieved polar effects caused by nonsense mutations in the phage genome or by chloramphenicol treatment of the host cells, and (iii) polarity was not caused by premature termination of transcription at the site of the polar mutation. RNA synthesis continued beyond the site to the first rhosensitive site.

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Section: Methodsmentioning

confidence: 99%

Bacteriophage phi X174-specific mRNA synthesis in cells deficient in termination factor rho activity

Imai¹

1981

J Virol

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“…Unlike other coliphages, such as T7, ϕX174 does not encode an RNA polymerase (RNAP), and as a result, is totally dependent on the E. coli host’s transcription system. The ϕX174 RNAP promoter locations were previously identified by a combination of binding assays (Chen et al, 1973) and hybridization mapping (Axelrod, 1976a, b), revealing three major promoters upstream of genes A, B, and D (Fig. 1A and Supplementary Table S1).…”

Section: Introductionmentioning

confidence: 97%

“…1A) (Hayashi et al, 1981; Hayashi et al, 1989). However, past work shows a potentially more complex picture where in vitro experiments detected a single Rho-dependent terminator in gene A (Axelrod, 1976a, b) and multiple Rho-dependent terminators in gene F (Axelrod, 1976a, b; Kapitza et al, 1979).…”

Section: Introductionmentioning

confidence: 99%

A high-resolution map of bacteriophage øX174 transcription

Logel

Jaschke

2020

Preprint

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Bacteriophage øX174 is a model virus for studies across the fields of structural biology, genetics, gut microbiome, and synthetic biology, but did not have a high-resolution transcriptome until this work. In this study we used next-generation sequencing to measure the RNA produced from øX174 while infecting its host E. coli C. We broadly confirm the past transcriptome model while revealing several interesting deviations from previous knowledge. Additionally, we measure the strength of canonical øX174 promoters and terminators and discover both a putative new promoter that may be activated by heat shock sigma factors, as well as rediscover a controversial Rho-dependent terminator. We also provide evidence for the first antisense transcription observed in the Microviridae family, identify two promoters that may be involved in generating this transcriptional activity, and discuss possible reasons why this RNA may be produced.

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“…Genes and numbering system are as described by Sanger et al (23). The filled triangles indicate the three known promoters (1,2,10,23); the open triangles indicate two other sites for which the evidence is incomplete (28). The bars represent the approximate location of termination sites observed by Hayashi et al (11), by Axelrod (1,2), by Smith and Sinsheimer (24), and by Kapitza et al (14,15), respectively.…”

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confidence: 99%

“…The four sites between nucleotides 5062 and 100 map in the region termed Tc (7,24). Sites 1389 and 1539 may correspond to the in vitro rho-de-pendent terminators T3a and T3b (1,2). The algorithm, based on a compilation of independent terminators, has previously been shown to also pick out some rho-dependent sites (3), which hints to some sequence similarity between dependent and independent terminators.…”

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confidence: 99%

Mapping of transcription terminators of bacteriophages phi X174 and G4 by sequence analysis

Brendel¹

1985

J Virol

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Transcription of bacteriophage φX174 in vitro: Analysis with restriction enzymes

Cited by 21 publications

References 11 publications

Bacteriophage phi X174-specific mRNA synthesis in cells deficient in termination factor rho activity

Bacteriophage phi X174-specific mRNA synthesis in cells deficient in termination factor rho activity

A high-resolution map of bacteriophage øX174 transcription

Mapping of transcription terminators of bacteriophages phi X174 and G4 by sequence analysis

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