Abortive infection of Escherichia coli with the bacteriophage f1: Cytoplasmic membrane proteins and the f1 DNA-gene 5 protein complex

Webster, Reginald P.; Cashman, Jay S.

doi:10.1016/s0042-6822(73)81005-0

Cited by 64 publications

(20 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Two of these proteins, pVII and pIX, are necessary for efficient particle assembly; the other two, pVI and pIII, are required for particle stability and phage infectivity (29,31). All five structural proteins are anchored in the inner membrane prior to incorporation into phage particles (4,6,35). Amino acid homologies and structural properties allowed us to identify putative equivalents of these five structural proteins on the B5 phage genome.…”

mentioning

confidence: 99%

Filamentous Phage Active on the Gram-Positive BacteriumPropionibacterium freudenreichii

et al. 2002

View full text Add to dashboard Cite

We present the first description of a single-stranded DNA filamentous phage able to replicate in a grampositive bacterium. Phage B5 infects Propionibacterium freudenreichii and has a genome consisting of 5,806 bases coding for 10 putative open reading frames. The organization of the genome is very similar to the organization of the genomes of filamentous phages active on gram-negative bacteria. The putative coat protein exhibits homology with the coat proteins of phages PH75 and Pf3 active on Thermus thermophilus and Pseudomonas aeruginosa, respectively. B5 is, therefore, evolutionarily related to the filamentous phages active on gram-negative bacteria.Filamentous phages are flexible rods that contain circular single-stranded DNA (ssDNA) coding for about 10 genes. They infect gram-negative bacteria by using a unique strategy of virion morphogenesis. Progeny particles are continually extruded or secreted across the bacterial membranes, causing neither lysis nor cell death (for reviews see references 22 and 23). This mode of assembly and excretion is related to bacterial processes responsible for the assembly of type 4 pili and for type II protein secretion (30). Many different filamentous phages have been characterized, and the Escherichia coli Ff phage family (M13, fd, and f1) is the best studied. Previous studies led to a better understanding of the role of these phages in bacterial evolution and the of spread of virulence factors. They also led to important applications in DNA sequencing and phage display techniques. Until now, there have been only infrequent reports of Ff-related phage active on gram-positive bacteria. A filamentous virus-like particle has been isolated from Clostridium acetobutylicum (17). However, the infectivity of this particle was not demonstrated, and its DNA was not characterized. More recently, five phages with filamentous morphology were isolated during a survey of phages active on Propionibacterium freudenreichii (9), a bacterium widely used in Swiss-type cheese manufacture to produce the characteristic flavor and eyes. In this work, we characterized one of these phages, a phage designated B5. This phage contains a ssDNA consisting of 5,806 bases and has a genome organization similar to the genome organization of other filamentous phages. This is the first description of an ssDNA phage genome that replicates in a gram-positive bacterium.Phage B5 was propagated on P. freudenreichii strain TIL18 in Yel medium (12) at 30°C and was enumerated by using the soft agar layer method (1). One-step growth experiments (1) revealed a latent period of about 11 h that corresponded to the generation time of the host. Phage B5 particles were filamentous, and the average size was 620 nm long by 12 nm wide (Fig. 1). Phage DNA, extracted as previously described (8), was resistant to all restriction enzymes but was degraded by S1 nuclease. These results suggested that like the genomes of all filamentous phages described so far, the phage B5 genome consists of ssDNA. Therefore, upon infection phage DNA ...

show abstract

mentioning

confidence: 99%

Filamentous Phage Active on the Gram-Positive BacteriumPropionibacterium freudenreichii

et al. 2002

View full text Add to dashboard Cite

show abstract

“…BL214, an RNase III-strain (14), was obtained from W. Studier. Bacteriophage f1, fi amber mutants, and media have been described (15). fl-Rlll, a gene V temperature-sensitive mutant, was received from P. Model.…”

mentioning

confidence: 99%

Bacteriophage f1 infection of Escherichia coli: identification and possible processing of f1-specific mRNAs in vivo.

Cashman¹,

Webster²

1979

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

ABSTRACT[3HJUracil-pulse-labeled RNA from Eschenichia coli infected with fl bacteriophage was fractionated on polyacrylamide gels containing urea. Eight phage-specific RNA species were present with approximate lengths ranging from 2100 to 400 nucleotides. The amount of the seven largest species was increased when the infected bacteria were incubated at 420C. When the RNA was isolated and used as message in an in vitro protein-synthesizing system, most of the RNA species appeared to direct the synthesis of the phage gene VIII protein.The six largest species also directed the synthesis of the phage gene V protein. Some of the labeled smaller RNA species increased in amount after addition of rifampicin, suggesting that they may have resulted from cleavage of larger RNA species. These particular smaller RNA species also were present in infected bacteria containing a mutant RNase III. The data are discussed in terms of the regulation of synthesis of the phagespecific proteins.The closely related filamentous bacteriophage f1, fd, and M13 contain a single-stranded DNA that codes for approximately eight genes (1, 2). Escherichia coli infected with these phage continue to grow and divide, while newly synthesized phage are extruded through the membrane. The identification of most phage products has been hampered by this continuation of host growth. However, products of the phage genes V and VIII (coat protein) are synthesized in large quantities and can be easily identified (3, 4). The two proteins are also the major products of coupled transcription-translation directed in vitro by replicative form DNA (5, 6). These findings suggest that there is some regulatory mechanism operating both in vivo and in vitro that allows this differential expression of phage products.Such a regulatory process may be operating partially at the level of transcription. Studies in vitro have shown that a small number of discrete transcripts are synthesized; initiation is from various promoters, while termination appears to occur at a unique site (7-11). Due to their location in the genome (12), the gene V and VIII regions are immediately proximal to the termination site; thus, most mRNAs in vitro would contain the coding information for these genes.In order to examine the basis for the high level of expression of the gene V and VIII proteins in vivo, it is necessary to characterize phage-specific mRNA found in infected bacteria. We report here the identification of a number of phage-specific mRNAs and show that several of them can direct the synthesis of the gene V and VIII proteins. Some of the species appear to correspond to those found in vitro, while others may have arisen as a result of processing from larger RNA molecules. These results are discussed in terms of the regulation of synthesis of the phage-specific proteins.MATERIALS AND METHODS E. coli K38, a nonsuppressing strain, and K37, a serine-inserting suppressing strain, were used (13). BL214, an RNase III-strain (14), was obtained from W. Studier. Bacteriophage f1, fi amber mutants...

show abstract

“…In the 140S This process resembles somewhat the final steps of phage assembly of the filamentous single-stranded DNA phages. Here a single-stranded DNA-gene 5 protein complex is found, which is converted into an intact phge with a complete stripping-off of the gene 5-protein (3,15). Possibly the Dprotein and the gene 5-protein have a similar function, i.e., protection of the nascent single-stranded DNA from duplication or degradation.…”

Section: Discussionmentioning

confidence: 99%

A DNA-Protein Complex Involved in Bacteriophage ϕX174 Particle Formation

Weisbeek

Sinsheimer

1974

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

A OX-specific DNA-protein complex has been isolated from 4X-infected cells. This complex contains infective circular single-stranded DNA, the proteins of the genes F, G, H, and J in the same proportions as in the phage particle, and, in addition, the gene D-protein.The D-protein makes up 12-22% of the protein part of the complex. The sedimentation value of the complex is about 140 S. In vitro, the complex can be converted to the normal 114S phage particle (with a concomitant loss of its D-protein) or to an uninfective 70S particle and a small amount of free single-stranded DNA. The fast sedimenting particle is not associated with membranes.The assembly of the mature OX174 phage particle seems to be closely linked to the synthesis of the viral single-stranded DNA (1). No condition is known where single-stranded DNA is formed without concomitant formation of phage particles. Under some conditions, a small amount of free single-stranded DNA has been found in infected cells (2), but always in combination with the synthesis of large amounts of phage.

show abstract

Abortive infection of Escherichia coli with the bacteriophage f1: Cytoplasmic membrane proteins and the f1 DNA-gene 5 protein complex

Cited by 64 publications

References 30 publications

Filamentous Phage Active on the Gram-Positive BacteriumPropionibacterium freudenreichii

Filamentous Phage Active on the Gram-Positive BacteriumPropionibacterium freudenreichii

Bacteriophage f1 infection of Escherichia coli: identification and possible processing of f1-specific mRNAs in vivo.

A DNA-Protein Complex Involved in Bacteriophage ϕX174 Particle Formation

Contact Info

Product

Resources

About