Isolation and characterization of bacteriophage T3/T7 hybrids and their use in studies on molecular basis of DNA-packaging specificity

Yamagishi, M; Fujisawa, Hisao; Minagawa, Teiichi

doi:10.1016/0042-6822(85)90290-9

Cited by 21 publications

(7 citation statements)

References 28 publications

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“…T3 and T7 package homologous DNA more efficiently than heterologous DNA. Gp19 is involved in the specificity for packaging the homologous DNA, and the DNA sequence responsible for recognition is located within 5% of the termini of the phage genomes [32]. Thus in the present T3/7 phage, the crossover in gene 17 can enhance adsorption to certain hosts, and that in gene 18.5 can assure that gene 19 and the right end terminal DNA retain the sequences of the T3 phage.…”

Section: Resultsmentioning

confidence: 99%

A T3 and T7 Recombinant Phage Acquires Efficient Adsorption and a Broader Host Range

Lin

Tseng

et al. 2012

PLoS ONE

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It is usually thought that bacteriophage T7 is female specific, while phage T3 can propagate on male and female Escherichia coli. We found that the growth patterns of phages T7M and T3 do not match the above characteristics, instead showing strain dependent male exclusion. Furthermore, a T3/7 hybrid phage exhibits a broader host range relative to that of T3, T7, as well as T7M, and is able to overcome the male exclusion. The T7M sequence closely resembles that of T3. T3/7 is essentially T3 based, but a DNA fragment containing part of the tail fiber gene 17 is replaced by the T7 sequence. T3 displays inferior adsorption to strains tested herein compared to T7. The T3 and T7 recombinant phage carries altered tail fibers and acquires better adsorption efficiency than T3. How phages T3 and T7 recombine was previously unclear. This study is the first to show that recombination can occur accurately within only 8 base-pair homology, where four-way junction structures are identified. Genomic recombination models based on endonuclease I cleavages at equivalent and nonequivalent sites followed by strand annealing are proposed. Retention of pseudo-palindromes can increase recombination frequency for reviving under stress.

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Section: Resultsmentioning

confidence: 99%

A T3 and T7 Recombinant Phage Acquires Efficient Adsorption and a Broader Host Range

Lin

Tseng

et al. 2012

PLoS ONE

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“…Subsequent extensive studies have supported this categorization. Procapsid binding components in the well-studied phages, other than phi29, include gpA in phage lambda (49), gp12 in phi21 (50), gp17 in T4 (51), gp19 in T3 and T7 (52,53). The DNA interacting components include gpNu1 in lambda (54,55), gp1 in phi21 (56), gp16 in T4 (57) and gp18 in T3/T7 (58,59).…”

Section: Discussionmentioning

confidence: 99%

Binding of pRNA to the N-terminal 14 amino acids of connector protein of bacteriophage phi29

Xiao

2005

Nucleic Acids Research

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During assembly, bacterial virus phi29 utilizes a motor to insert genomic DNA into a preformed protein shell called the procapsid. The motor contains one twelve-subunit connector with a 3.6 nm central channel for DNA transportation, six viral-encoded RNA (packaging RNA or pRNA) and a protein, gp16, with unknown stoichiometry. Recent DNA-packaging models proposed that the 5-fold procapsid vertexes and 12-fold connector (or the hexameric pRNA ring) represented a symmetry mismatch enabling production of a force to drive a rotation motor to translocate and compress DNA. There was a discrepancy regarding the location of the foothold for the pRNA. One model [C. Chen and P. Guo (1997) J. Virol., 71, 3864-3871] suggested that the foothold for pRNA was the connector and that the pRNA-connector complex was part of the rotor. However, one other model suggested that the foothold for pRNA was the 5-fold vertex of the capsid protein and that pRNA was the stator. To elucidate the mechanism of phi29 DNA packaging, it is critical to confirm whether pRNA binds to the 5-fold vertex of the capsid protein or to the 12-fold symmetrical connector. Here, we used both purified connector and purified procapsid for binding studies with in vitro transcribed pRNA. Specific binding of pRNA to the connector in the procapsid was found by photoaffinity crosslinking. Removal of the N-terminal 14 amino acids of the gp10 protein by proteolytic cleavage resulted in undetectable binding of pRNA to either the connector or the procapsid, as investigated by agarose gel electrophoresis, SDS-PAGE, sucrose gradient sedimentation and N-terminal peptide sequencing. It is therefore concluded that pRNA bound to the 12-fold symmetrical connector to form a pRNA-connector complex and that the foothold for pRNA is the connector but not the capsid protein.

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“…One crucial function of the connector is to serve as a nucleation point for the assembly of motor components. The binding of a large subunit of DNA packaging enzymes (see below) to the connector has been reported, including gpA in phage λ ( Catalano et al ., 1995 ), gp12 in phi21 ( Feiss et al ., 1985 ), gp17 in T4 ( Rao and Black, 1988 ) and gp19 in T3 and T7 ( Yamagishi et al ., 1985 ; Morita et al ., 1993 ), and both gp16 and pRNA of phi29 ( Guo et al ., 1987b ; Chen and Guo, 1997 ; Garver and Guo, 2000 ; Xiao, et al ., 2005 ; Lee and Guo, 2006 ). For phage λ, it has been found that the target component of gpA binding is the connector protein, because double mutation studies with the gpA and connector revealed that a connector mutation can suppress a gpA mutation that hinders gpA binding to procapsid ( Yeo and Feiss, 1995 ).…”

Section: Essential Components Of Viral Dna Packaging Motormentioning

confidence: 99%

“…The larger component containing the conserved ATP‐binding motif is involved in procapsid binding, while the smaller component binds DNA (Guo et al ., 1987a), as confirmed by subsequent studies in many viral systems. Procapsid binding components in well‐studied phages include gpA in λ (Weisberg et al ., 1979; Yeo and Feiss, 1995), gp12 in phi21 (Feiss et al ., 1985), gp17 in T4 (Rao and Black, 1988), gp19 in T3/T7 (Yamagishi et al ., 1985), g2p in SPP1 (Gual et al ., 2000), g2p in SF6 (Chai et al ., 1992), p9 of PRD1 (Stromsten et al ., 2005), gp2 in P22 (Casjens et al ., 1992) and gpP in P2/P4 (Rishovd et al ., 1998). The DNA interacting components include gpNu1 in λ, gp1 in phi21, gp16 in T4, gp18 in T3/T7, g1p in SPP1, g1p in SF6 and gp3 in P22.…”

Section: Essential Components Of Viral Dna Packaging Motormentioning

confidence: 99%

Viral nanomotors for packaging of dsDNA and dsRNA

Guo

Lee

2007

Molecular Microbiology

104

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SummaryWhile capsid proteins are assembled around singlestranded genomic DNA or RNA in rod-shaped viruses, the lengthy double-stranded genome of other viruses is packaged forcefully within a preformed protein shell. This entropically unfavourable DNA or RNA packaging is accomplished by an ATP-driven viral nanomotor, which is mainly composed of two components, the oligomerized channel and the packaging enzymes. This intriguing DNA or RNA packaging process has provoked interest among virologists, bacteriologists, biochemists, biophysicists, chemists, structural biologists and computational scientists alike, especially those interested in nanotechnology, nanomedicine, AAA+ family proteins, energy conversion, cell membrane transport, DNA or RNA replication and antiviral therapy. This review mainly focuses on the motors of doublestranded DNA viruses, but double-stranded RNA viral motors are also discussed due to interesting similarities. The novel and ingenious configuration of these nanomotors has inspired the development of biomimetics for nanodevices. Advances in structural and functional studies have increased our understanding of the molecular basis of biological movement to the point where we can begin thinking about possible applications of the viral DNA packaging motor in nanotechnology and medical applications.

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Isolation and characterization of bacteriophage T3/T7 hybrids and their use in studies on molecular basis of DNA-packaging specificity

Cited by 21 publications

References 28 publications

A T3 and T7 Recombinant Phage Acquires Efficient Adsorption and a Broader Host Range

A T3 and T7 Recombinant Phage Acquires Efficient Adsorption and a Broader Host Range

Binding of pRNA to the N-terminal 14 amino acids of connector protein of bacteriophage phi29

Viral nanomotors for packaging of dsDNA and dsRNA

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