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PubMed1969

Identification of closely linked loci controlling ultraviolet sensitivity and refractivity to colicin E2 in Escherichia coli.

Mutants (phenotypic symbol Ref-II) refractory to colicin E2 have been isolated in several strains of Escherichia coli K-12, and a refII locus has been mapped 1 to 2 min counter clockwise to thr. A small number of Ref-II mutants are also ultraviolet (UV)-sensitive and the uv(s) locus in one such strain has been mapped close to the refII locus near thr. The Ref-II mutation alone does not affect recombinant formation in F(-) strains, but the Ref-II, UV(s) strains behave in many respects like Rec(-) mutants, giving reduced recombination frequencies in crosses with male strains. It is suggested that the refII and uv(s) loci correspond to closely linked if not identical genes, concerned in some way in the activity of one or more deoxyribonucleases, and that the Ref-II, UV(s) mutants arise as the pleiotropic expression of a single gene or of a deletion or polar mutation affecting linked genes.

Chromosome Mapping↗
PubMed1969

Carbohydrate accumulation and metabolism in Escherichia coli: the close linkage and chromosomal location of ctr mutations.

Six pleiotropic ctr mutations of Escherichia coli, affecting the ability to utilize 10 carbohydrates, were found to be closely linked to one another and to the mutation of strain MM6 causing lack of enzyme 1 of the phosphotransferase system. These mutations are located at 46 to 47 min on the E. coli map. Preliminary biochemical evidence indicates that the ctr mutants also lack enzyme 1, although they have a different phenotype from MM6.

Carbohydrate Metabolism↗
PubMed1969

Chromosomal location of genes regulating resistance to bacteriophage in Bacillus subtilis.

Many of the viruses which infect Bacillus subtilis require glucosylated polyglycerol teichoic acid for adsorption. These mutants can be divided into three classes on the basis of enzymatic defects and growth on galactose-minimal medium. Transduction with phage PBS1 reveals that two of these, gtaA and gtaB, are linked to hisA1, whereas the gtaC locus is linked to argC. Analysis by deoxyribonucleic acid-mediated transformation indicates that these loci exist in a cluster between the hisA1 and argC4 loci. Anomalies in mapping in the group II region of the chromosome exist. The basis of these anomalies is discussed.

Adsorption↗
PubMed1969

Linked transformation of bacterial and prophage markers in Bacillus subtilis 168 lysogenic for bacteriophage phi 105.

Level of competence reached by Bacillus subtilis 168 lysogenic for temperate phage phi 105 was reduced compared to that reached by nonlysogenic cells. This effect was probably related to an alteration of the bacterial surface. Deoxyribonucleic acid extracted from phi 105 lysogenic bacteria was used to transform other lysogenic bacteria. About 25% linkage was found between the bacterial phe-1 marker and prophage marker ts N15. The order of a few prophage markers relative to phe-1 was established in three-factor crosses. The usefulness of this system for a study of the linkage between an integrated prophage genome and that of its host was discussed.

Bacillus subtilis↗
PubMed1969

Leakiness of pleiotropic maltose-negative, bacteriophage lambda-resistant mutants of Escherichi coli K-12.

Cultures of Escherichia coli K-12 malA(-)lambda(r)gal(-) can be transduced to gal(+) by bacteriophage lambdadg because of leakiness of the lambda(r) phenotype. The efficiency of such transduction is about 10(-5) that of transduction of mal(+)lambda(s) bacteria. Leaky cells (lambda(s)phenocopies) adsorb only very few phage particles, and many transductants, therefore, are defective heterogenotes or show integration of the gal(+) gene, which is unaccompanied by lysogenization.

Bacteriophages↗
PubMed1969

Variations in genetic recombination due to amber mutations in T4D bacteriophage.

Recombination experiments were performed to assess the affect of amber mutations in 12 genes of T4D bacteriophage on genetic recombination. Crosses were performed in various suppressor-containing bacterial hosts to permit the production of progeny phage. Amber mutations in genes 32, 46, and 47 caused decreased recombination, amber mutations in genes 30, 41, 42, 43, 56, 61, and 62 caused increased recombination, whereas mutations in genes 63 and 37 showed no demonstrable effect on recombination.

Coliphages↗
PubMed1969

Nature of Escherichia coli B(P1) yielder cells at the time of infection with restricted T1.

In the infection of Escherichia coli B(P1) with restricted T1, it was shown that yielder cells consist of both special and nonspecial cells. Special or predetermined yielders occurred only among the earliest yielders. In most instances, yielder-cell formation was most easily explained by assuming that the first step was a chance escape of the restricted phage DNA from the degrading enzyme of the restricting cell.

Coliphages↗
PubMed1969

In vitro transformation by adenovirus-simiam virus 40 hybrid viruses. IV. Properties of clones isolated from cell lines transformed by adenovirus 2-simiam virus 40 and adenovirus 12-simiam virus 40 transcapsidant hybird viruses.

Clones were isolated from hamster cells transformed by the adenovirus 2-SV40 and adenovirus 12-SV40 transcapsidant hybrid viruses. The clones were characterized with respect to their cytomorphology, virus and antigen content, and the histomorphology of tumors induced by transplantation of the clonal sublines to hamsters. Three different cellular and colonial morphologies were observed. Clones with an SV40 morphology gave rise to tumors predominantly with an SV40 histology, whereas clones with an adenovirus morphology produced typical adenovirus tumors upon transplantation of the transformed cells. Clones which had features of both SV40 and adenovirus transformed cells gave rise to "intermediate" and adenovirus tumors. The results indicate that multiple events occur during transformation and tumorigenesis by the transcapsidant virus populations and provide an explanation for the multiplicity of findings which have been reported with these virus populations.

Adenoviridae↗
PubMed1970

Characterization of inducible bacteriophages in Bacillus licheniformis.

Two morphologically distinct and physically separable defective phages have been found in Bacillus licheniformis NRS 243 after induction by mitomycin C. One of them (PBLB) is similar to the defective phage PBSX of B. subtilis, which has a density of 1.373 g/cm(3) in CsCl and a sedimentation coefficient of 160S. PBLB incorporates into its head mainly bacterial deoxyribonucleic acid (DNA) which has a sedimentation coefficient of 22S and a buoyant density in CsCl of 1.706 g/cm(3). The other phage (PBLA) has a morphology similar to the temperate phage phi105 of B. subtilis; the head diameter is about 66 nm, and it possesses a long and noncontractile tail. PBLA has a density of 1.484 g/cm(3) in CsCl and the phage-specific DNA, which is exclusively synthesized after induction by mitomycin C, has a density of 1.701 g/cm(3). PBLA DNA is double-stranded and has a sedimentation coefficient of 36S, corresponding to a molecular weight of 34 x 10(6) to 35 x 10(6) daltons. The phage DNA has one interruption per single strand, giving single-stranded segments with molecular weights of 13 x 10(6) and 4 x 10(6) daltons. Common sequences between the two phage DNA species and with their host DNA have been demonstrated by DNA-DNA hybridization studies. Both phage particles kill sensitive bacteria. However, all attempts thus far to find an indicator strain to support plaque formation have been unsuccessful.

Bacillus↗
PubMed1970

Deoxyribonucleic acid replication in simian virus 40-infected cells. II. Detection and characterization of simian virus 40 pseudovirions.

Purified simian virus 40 (SV40) virions, grown in primary African green monkey kidney cells labeled prior to infection with (3)H-thymidine, contain a variable quantity of (3)H-labeled deoxyribonucleic acid (DNA). This DNA is resistant to deoxyribonuclease, sediments at 250S, and is enclosed in a particle that can be precipitated with SV40-specific antiserum. DNA-DNA hybridization experiments demonstrate that this (3)H-labeled component in purified SV40 virions is cellular DNA. When this (3)H-labeled DNA is released from purified virus with sodium dodecyl sulfate, it has an average sedimentation constant of 14S. Sedimentation through neutral and alkaline sucrose gradients shows that this 14S DNA is composed of a collection of different sizes of DNA molecules that sediment between 11 and 15S. As a result of this size heterogeneity, SV40 virions containing cellular DNA (pseudovirions) have a variable DNA to capsid protein ratio and exhibit a spectrum of buoyant densities in a CsCl equilibrium gradient. Pseudovirions are enriched, relative to true virions, on the lighter density side of infectious SV40 virus banded to equilibrium in a CsCl gradient. Little or no cellular DNA was found in purified SV40 virus preparations grown in BSC-1 or CV-1 cells.

Animals↗
PubMed1970

Transfer ribonucleic acid in KB cells infected with adenovirus type 2.

Populations of transfer ribonucleic acid (tRNA) extracted from control and type 2 adenovirus (Ad2)-infected KB cells were compared. No consistent differences in acceptor activity for 11 amino acids were observed. Comparison of methylated albumin-kieselguhr (MAK) elution profiles of arginyl-tRNA from control and infected cells revealed a minor modification in that the proportion of arginyl-tRNA eluting at high salt concentration was somewhat greater in infected cells. No similar differences were observed in MAK elution profiles of aspartyl-, isoleucyl-, leucyl-, phenylalanyl-, seryl-, tyrosyl-, and valyl-tRNA. Hybridization of 4S RNA from infected cells labeled by incorporation of (3)H-uridine with Ad2 deoxyribonucleic acid revealed the presence of a complementary species of RNA in this preparation. Hybridization of (3)H-arginyl-tRNA and of (3)H-aminoacyl-tRNA labeled by charging with (3)H-arginine or a (3)H-mixture of amino acids, respectively, failed to detect the presence of virus-specific tRNA in Ad2-infected cells.

Adenoviridae↗
PubMed1970

Deoxyribonucleic acid replication in simian virus 40-infected cells. 3. Comparison of simian virus 40 lytic infection in three different monkey kidney cell lines.

A comparative study of simian virus 40 (SV40) lytic infection in three different monkey cell lines is described. The results demonstrate that viral deoxyribonucleic acid (DNA) synthesis and infectious virus production begin some 10 to 20 hr earlier in CV-1 cells and primary African green monkey kidney (AGMK) cells than in BSC-1 cells. Induction of cellular DNA synthesis by SV40 was observed in CV-1 and AGMK cells but not with BSC-1 cells. Excision of large molecular weight cellular DNA to smaller fragments was easily detectable late in infection of AGMK cells. Little or no excision was observed at comparable times after infection of CV-1 and BSC-1 cells. The different kinds of responses of these three monkey cell lines during SV40 lytic infection suggest the involvement of cellular functions in the virus-directed induction of cellular DNA synthesis and the excision of this DNA from the genome.

Animals↗
PubMed1970

Virus-specific deoxyribonucleic acid in simian virus 40-exposed hamster cells: correlation with S and T antigens.

Several homologous hamster embryonic cell lines, transformed in association with simian virus (SV) 40 infection, were examined for the presence of deoxyribonucleic acid (DNA) complementary to SV40 ribonucleic acid (RNA) made in vitro. The methods employed permitted the detection of 10(-5) mug of viral DNA in 100 mug of cellular DNA, corresponding to one-fifth of an SV40 DNA molecule per cell. Those lines which contained both the SV40 surface (S) and tumor (T) antigens also contained DNA complementary to SV40 RNA synthesized in vitro. In contrast, neither of two lines which contained S, but not T, antigen contained detectable DNA complementary to SV40 RNA. These findings suggest that the production of S antigen does not depend upon the persistence of SV40 DNA in transformed cells.

Animals↗
PubMed1970

Inhibition of host protein synthesis during infection of Escherichi coli by bacteriophage T4. 3. Inhibition by ghosts.

Deoxyribonucleic acid (DNA)-less T2 "ghosts" were prepared by osmotic shock and purified by KBr density gradient centrifugation. Escherichia coli B was treated with these ghosts in inorganic salts-glycerol medium to see which features of phage infection could be elicited by ghosts. At a multiplicity that was just sufficient to block induction of beta-galactosidase (EC 3.2.1.23), 89% of the bacteria were killed and the rates of ribonucleic acid (RNA) and DNA synthesis were about 10 to 15% of normal. However, protein synthesis was almost completely blocked but resumed after 30 min. During this period, it was possible to induce messenger RNA (mRNA) from the lactose operon, although this mRNA could not be translated into active beta-galactosidase. These results suggest to us that the viable cells surviving ghost infection synthesize nucleic acids at close to a normal rate but are temporarily blocked in protein synthesis. The continued formation of untranslated host mRNA mimics the pattern of bacterial synthesis just after whole-phage infection, and is consistent with the interpretation that the immediate block in the initiation of host translation by these viruses is due to their attachment.

Bacterial Proteins↗
PubMed1971

Size and composition of Marek's disease virus deoxyribonucleic acid.

Deoxyribonucleic acid (DNA) extracted from purified nucleocapsids of Marek's disease herpesvirus (MDV) was cosedimented with T4 and with herpes simplex virus (HSV) DNA in neutral sucrose density gradients and with T4 DNA in alkaline sucrose density gradients. These experiments indicated that the intact MDV DNA had a sedimentation constant of 56S corresponding to a molecular weight of 1.2 x 10(8) daltons. In the alkaline gradients, the largest and most prominent band contains a DNA sedimenting at 70S corresponding to 6.0 x 10(7) daltons in molecular weight. The DNA is therefore double-stranded and not cross-linked. Isopycnic sedimentation of the MDV DNA molecules with SPO1, Micrococcus lysodeikticus, and HSV DNA gave a density of 1.705 g/cm(3) corresponding to 46 guanine plus cytosine moles per cent. Lastly, in hybridization tests the DNA hybridized with RNA of infected cells but not with that of uninfected cells supporting the conclusion that it is viral.

Alpharetrovirus↗
PubMed1971

Effect of rifamycins and related antibiotics on the deoxyribonucleic acid-dependent ribonucleic acid polymerase of vaccinia virus particles.

A number of compounds related to rifampin which act as expected in the Escherichia coli system have been tested for their ability to inhibit the vaccinia particle deoxyribonucleic acid-dependent ribonucleic acid (RNA) polymerase in vitro. Some compounds are inactive even at concentrations of 500 mug/ml, others are able to produce partial inhibition, and others strongly inhibit the enzyme activity at 150 mug/ml or less. The inhibition, where present, operates immediately but appears to be at least partially reversible. At least one compound which is without effect against bacterial RNA polymerase is a potent inhibitor of the viral RNA polymerase. As the enzyme activity of rifampin-resistant mutants of vaccinia virus is inhibited to the same extent as that of the wild type, the observed in vitro effect on vaccinia virus RNA polymerase is not identical with the in vivo effect specifically directed against a vaccinia-specified protein.

Agar↗
PubMed1971

Ribonucleic acid bacteriophage release: requirement for host-controlled protein synthesis.

The release of the ribonucleic acid (RNA)-containing phage MS2 from Escherichia coli is accompanied by cellular lysis at 37 C, whereas at 30 C phage are released from intact cells. Chloramphenicol or rifampin prevents the release of progeny phage particles at both temperatures. Neither drug causes an immediate cessation of phage release and after inhibition of protein synthesis by chloramphenicol phage release proceeds for about 17 min at 37 C and about 35 min at 30 C. Rifampin does not inhibit phage release from mutant cells possessing a rifampin-resistant deoxyribonucleic acid-dependent RNA polymerase. The results indicate that a short-lived host-controlled protein(s) is essential for the release of RNA phage particles at both temperatures.

Bacterial Proteins↗
PubMed1972

Genetic and physiological studies of bacteriophage T5. 3. Patterns of deoxyribonucleic acid synthesis induced by mutants of T5 and the identification of genes influencing the appearance of phage-induced dihydrofolate reductase and deoxyribonuclease.

Patterns of deoxyribonucleic acid (DNA) metabolism in nonpermissive cells infected with amber mutants representing 29 genes of T5 are reported. A group of 7 contiguous genes are essential for the synthesis of phage DNA, whereas 20 other genes, when defective, permit varying degrees of phage DNA synthesis. Two further genes are essential for complete transfer of phage DNA to host cells, and therefore indirectly do not permit the synthesis of phage DNA. The structural genes for an early T5 deoxyribonuclease and for T5 DNA polymerase, as well as a gene that affects the synthesis of dihydrofolate reductase, have been identified in the genetic map of T5.

Carbon Isotopes↗