Synthesis of Bacteriophage φ29 Proteins in
            <i>Bacillus subtilis</i>

Pène, Jacques J.; Murr, Peter; Barrow-Carraway, Joyce

doi:10.1128/jvi.12.1.61-67.1973

Cited by 26 publications

(14 citation statements)

References 8 publications

(6 reference statements)

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“…The genome is a linear, nonpermuted duplex DNA of 12 megadaltons (Mdal), with protein (gp3) covalently bound to the 5' termini (1,2,24,30,31,49,55). Eighteen cistrons have been mapped, and 25 virus-specific proteins, reflecting more than 90% of the coding capacity of the genome, have been identified (5,13,25,38,39,45).…”

mentioning

confidence: 99%

Morphogenesis of bacteriophage phi 29 of Bacillus subtilis: oriented and quantized in vitro packaging of DNA protein gp3

Bjornsti¹,

Reilly²,

Anderson³

1983

J Virol

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The assembly of phage 4)29 occurs by a single pathway, and the DNA protein (DNA-gp3) of "packaging intermediates" can be obtained after DNase I interruption of in vitro complementation. A broad spectrum of DNA molecules of variable length was isolated from DNase I-treated proheads. Restriction endonuclease EcoRI digestion and electrophoretic analysis of these DNA molecules suggested that DNA-gp3 packaging was oriented with respect to the physical map and was a complex process. Proteinase K-treated exogenous DNA was not packaged. When exogenous DNA-gp3 was predigested with the restriction endonucleases BstEII, EcoRI, HpaI, and HpaII, the left-end fragments, ranging in size from 8 to 0.9 megadaltons, were selectively and efficiently packaged. During in vivo and in vitro assembly, DNA-gp3 is packaged into proheads, the "core-scaffolding" protein gp7 exits from the particles, and the DNA-filled heads assume the angular morphology of phage 4)29. The packaging of a 4.1-megadalton DNA-gp3 left-end fragment (one third of the genome) resulted in the exit of gp7 and the transition to angularity. MATERIALS AND METHODS Chemicals and isotopes. Spermidine (A grade) and DNase I (bovine pancreatic) were obtained from Calbiochem-Behring Corp., San Diego, Calif. ATP (equine muscle) and egg white lysozyme (3 x recrystallized) were purchased from Sigma Chemical Co., St. Louis, Mo. ATP was dissolved in TMS buffer (0.05 M Tris-hydrochloride [pH 7.8], 0.01 M MgCl2, 0.1 M NaCI) and neutralized with 1.5 N ammonium hydroxide to give a stock solution of 20 mM (pH 7.8). Proteinase K (E. Merck, Darmstadt, West Germany) was dissolved in water. High-gelling-temperature agarose was from FMC Corp., Rockland, Maine, and 2mercaptoethanol was obtained from Eastman Chemical Co., Rochester, N.Y. The restriction endonucleases EcoRI, BstEIl, Haelll, and Hpall were purchased from Bethesda Research Labs, Rockville, Md., and Hpal was from Boehringer-Mannheim Biochemical, Indianapolis, Ind. [3H]thymidine (no. NET 0272; 50 Ci/mmol) and the "C-labeled amino acid mixture (no. NEC 445E; about 200 mCi/ mmol) were from New England Nuclear Corp., Boston, Mass. Phage and bacteria. Phage 4)29 and previously described mutants were used (39, 47). Recombinants were constructed with mutant susl4(1241), a mutation that results in the delayed-lysis phenotype (42). The properties of the permissive host B. subtilis sup44 (40) and the nonpermissive host B. siubtilis spoA12 (48) have been described previously.

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mentioning

confidence: 99%

Morphogenesis of bacteriophage phi 29 of Bacillus subtilis: oriented and quantized in vitro packaging of DNA protein gp3

Bjornsti¹,

Reilly²,

Anderson³

1983

J Virol

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“…Notably, upregulation of host38's translation genes was also middle/middle late [30], thus suggesting that phage phi38:1 translation in this host is mainly timed with the activation of host translation as occurs with phi18:3 on host18. Given that phage protein translation is expected to closely follow transcription based on lessons from phages infecting (efficiently) well-known systems (e.g., Bacillus phage phi29 synthesizes >60% of its proteins before middle transcription [77]), the inability of phi38:1 to synchronize translation with transcription in host18 may contribute to this inefficient infection.…”

Section: Absence Of Transcriptional-translational Synchronization In mentioning

confidence: 99%

Multiple mechanisms drive phage infection efficiency in nearly identical hosts

et al. 2018

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Phage-host interactions are critical to ecology, evolution, and biotechnology. Central to those is infection efficiency, which remains poorly understood, particularly in nature. Here we apply genome-wide transcriptomics and proteomics to investigate infection efficiency in nature's own experiment: two nearly identical (genetically and physiologically) Bacteroidetes bacterial strains (host18 and host38) that are genetically intractable, but environmentally important, where phage infection efficiency varies. On host18, specialist phage phi18:3 infects efficiently, whereas generalist phi38:1 infects inefficiently. On host38, only phi38:1 infects, and efficiently. Overall, phi18:3 globally repressed host18's transcriptome and proteome, expressed genes that likely evaded host restriction/modification (R/M) defenses and controlled its metabolism, and synchronized phage transcription with translation. In contrast, phi38:1 failed to repress host18's transcriptome and proteome, did not evade host R/M defenses or express genes for metabolism control, did not synchronize transcripts with proteins and its protein abundances were likely targeted by host proteases. However, on host38, phi38:1 globally repressed host transcriptome and proteome, synchronized phage transcription with translation, and infected host38 efficiently. Together these findings reveal multiple infection inefficiencies. While this contrasts the single mechanisms often revealed in laboratory mutant studies, it likely better reflects the phage-host interaction dynamics that occur in nature.

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“…imum number of complementation groups reported by an one laboratory is 17 (199); thus even if all cistrons reported by other groups are redundant, this appears to provide a minimum number of polypeptides whose synthesis is directed by 029 DNA. In UV-irradiated B. subtilis infected with 029, up to 23 phage-induced proteins have been separated by gel electrophoresis (48,122,195,227). It is possible, however, that some of the proteins identified on gels may be precursor proteins which are cleaved to generate lower-molecular-weight products which are observed on the same gels (7).…”

Section: Sp82gmentioning

confidence: 99%

“…Studies of phage-directed protein synthesis have been carried out by infecting UV-irradiated B. subtilis with wild-type or mutant 429 in the presence of radioactive amino acids, and subsequently separating the isolated proteins on polyacrylamide gels. Using this technique Carrascosa et al (47) detected 12 nonstructural proteins in addition to seven structural components; Pene and co-workers (227) found seven structural proteins and as many as 10 early and four late nonstructural polypeptides. Moreover, this latter group found that synthesis of six of the early nonstructural proteins ceased late in infection, and they suggest these proteins may be related to the early 750,000-molecularweight RNA transcript which is shut off at the time DNA synthesis is initiated.…”

Section: Development Of 4o29 429 Infection Has Little Effect On Host mentioning

confidence: 99%