The self-assembly of RNA free protein subunits from bacteriophage MS-2

Rohrmann, George F.; Krueger, Robert G.

doi:10.1016/0006-291x(70)90728-x

Cited by 15 publications

(10 citation statements)

References 11 publications

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“…Encapsulation is based on the specific interaction between the RNA hairpin operator, i.e., pac site, and the MS2 CP, leading to spontaneous assembly of the viral shell and packaging the molecules around them. It has been demonstrated that (Rohrmann and Krueger 1970;Stockley et al 1994;Witherell et al 1991) the conserved bases of the MS2 operator are the A-4 and A-7 positions of the single-stranded loop, the −5 position must be a pyrimidine for tight binding, and the mismatched base of −10 position can be a purine (Fig. 1c).…”

Section: Discussionmentioning

confidence: 99%

A novel method to produce armored double-stranded DNA by encapsulation of MS2 viral capsids

Zhang

Sun

Chang

et al. 2015

Appl Microbiol Biotechnol

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With the rapid development of molecular diagnostic techniques, there is a growing need for quality controls and standards with favorable properties to monitor the entire detection process. In this study, we describe a novel method to produce armored hepatitis B virus (HBV) and human papillomavirus (HPV) DNA for use in nucleic acid tests, which was confirmed to be stable, homogeneous, noninfectious, nuclease resistant, and safe for shipping. We demonstrated that MS2 bacteriophage could successfully package double-stranded DNA of 1.3-, 3-, 3.5-, and 6.5-kb length into viral capsids with high reassembly efficiency. This is the first application of RNA bacteriophage MS2 as a platform to encapsulate double-stranded DNA, forming virus-like particles (VLPs) which were indistinguishable from native MS2 capsids in size and morphology. Moreover, by analyzing the interaction mechanism of pac site and the MS2 coat protein (CP), we found that in addition to the recognized initiation signal TR-RNA, TR-DNA can also trigger spontaneous reassembly of CP dimers, providing a more convenient and feasible method of assembly. In conclusion, this straightforward and reliable manufacturing approach makes armored DNA an ideal control and standard for use in clinical laboratory tests and diagnostics, possessing prospects for broad application, especially providing a new platform for the production of quality controls for DNA viruses.

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Section: Discussionmentioning

confidence: 99%

A novel method to produce armored double-stranded DNA by encapsulation of MS2 viral capsids

Zhang

Sun

Chang

et al. 2015

Appl Microbiol Biotechnol

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“…The expression construct does not include the MS2 replicase translational operator which is normally located 3' to the coat protein gene and is believed to function as the assembly initiation sequence (Beckett et al, 1988). The formation of empty capsids upon coat protein expression is therefore accounted for (Rohrmann & Krueger, 1970). Soluble forms of the chimeric capsids are easily purified, but even insoluble forms can be recovered by a simple cycle of disassembly and reassembly in the presence of urea.…”

Section: Discussionmentioning

confidence: 99%

“…The system has a number of advantages over the filamentous bacteriophage alternatives. The MS2 coat protein is capable of facile self-assembly in the absence of nucleic acid (Rohrmann & Krueger, 1970) unlike the filamentous phages in which assembly is concomitant with encapsidation of nucleic acid (Model & Russel, 1988). Furthermore the filamentous coat protein must undergo post-translational processing and membrane insertion before assembly occurs whereas the MS2 protein is unprocessed.…”

Section: Discussionmentioning

confidence: 99%

Multiple presentation of foreign peptides on the surface of an RNA-free spherical bacteriophage capsid

Mastico

Talbot²,

Stockley³

1993

Journal of General Virology

136

120

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We have produced a plasmid expression vector for the coat protein of RNA bacteriophage MS2. The vector has been modified to introduce a unique Kpni restriction site within the coat protein gene at a site corresponding to the most radially distant feature of the bacteriophage capsid, namely the top of the N-terminal fl-hairpin (between residues 15 and 16). Insertion of DNA oligonucleotides at this site allows the production of chimeric MS2 coat proteins having foreign peptide sequences expressed as the central part of the hairpin. We have produced chimeras with a number of different peptide sequences (up to 24 amino acids in length) chosen because of their known antigenic properties. The chimeric coat proteins self-assemble into largely RNAfree phage-like capsids in Escherichia coli and can be easily disassembled and reassembled in vitro. Such peptide-presenting particles may have a number of biotechnological applications, including use as a costeffective, synthetic vaccine. We have tested the antigenicity of one such construct in vivo in mice and have shown that these particles are immunogenic and that antibody titres against the inserted peptide epitope can be obtained.

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“…2. MS-2 coat protein prepared by acetic acid treatment of infectious virus contained less than 1 % of the RNA of the H particles (23) and had a density of 1.33 g/cm3 (Table 1). Phage coat protein dissolved in 66% acetic acid was shown to contain a homogeneous population of 15,000molecular weight subunits in the model E analytical ultracentrifuge (23). When coat protein in 0.01 M acetic acid was analyzed by using polyacrylamide disc electrophoresis, two molecular species were evident with the larger one predominating (23).…”

mentioning

confidence: 99%

“…The coat protein of phage MS-2 and that of M-12 have identical amino acid sequences and differ by two different amino acid exchanges at the same position from the coat proteins of phages R-17 and f2 (4, 12, 30). The coat protein of the RNA coliphages is capable of aggregating under proper experimental conditions to form a particle that is antigenically identical to capsids but of higher molecular weight (9,10,23).…”

mentioning

confidence: 99%

Physical, Biochemical, and Immunological Properties of Coliphage MS-2 Particles

Rohrmann

Krueger

1970

J Virol

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H (heavy) and L (light) MS-2 particles differ in density, absorption spectrum, and infectivity. Studies on their sedimentation, ribonucleic acid (RNA) content and infectivity, appearance under the electron microscope, ribonuclease sensitivity, and A-protein content failed to demonstrate any difference between the two particle types. Studies on the size, RNA content, and density of the capsid and two smaller coat protein components were also conducted. The antigenic relatedness of five different viral and subviral particles of MS-2 were studied by using immunodiffusion and neutralization. Capsids and the H and L viral particles were shown to be antigenically related, whereas the coat protein monomers and dimers were shown to be unrelated to the higher-molecular-weight particles.

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The self-assembly of RNA free protein subunits from bacteriophage MS-2

Cited by 15 publications

References 11 publications

A novel method to produce armored double-stranded DNA by encapsulation of MS2 viral capsids

A novel method to produce armored double-stranded DNA by encapsulation of MS2 viral capsids

Multiple presentation of foreign peptides on the surface of an RNA-free spherical bacteriophage capsid

Physical, Biochemical, and Immunological Properties of Coliphage MS-2 Particles

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