MS2 phage

Hasegawa M., Yasunaga T., Miyata T. (1979). Secondary structure of MS2 phage RNA and bias in code word usage. Nucleic Acids Res. 7 2073-2079. 

Weissmann, C. and Borst, P. (1963) Double-stranded ribonucleic acid formation in vitro by MS2 phage-induced RNA synthetase. Science, 142, 1188-1191. 

Fig. 32. Percentage of the input P from labeled MS2 phage into a form resistant to ribonuclease in a lysate of cells as a function of time after removal of actinomycin (Kelly et al., 1965).

Introducing indicator organisms into the membrane feed water such as MS2 phage. 

Differential inhibitory effects of antibiotics on the synthesis of membrane and cytoplasmic proteins may be due to differences in the ribosomes themselves. Differences in the ribosomes of E, coli may result from the protein components or the RNA components. In fact, some ribo-somal proteins have been shown to be present in only a fraction of ribosomes, resulting in a heterogeneity of ribosomes. The fact that there are several nonlinked ribosomal RNA genes in an E. coli genome may reflect the differentiation of ribosomes. This differentiation of ribosomes may be the direct cause of the different sensitivities of ribosomes to antibiotics. Alternatively, the different sensitivities of ribosomes to antibiotics could be caused indirectly as a result of compartmentalization of ribosomes inside the cell. Existence of a subpopulation of ribosomes with different sensitivities has been suggested for MS2 phage protein synthesis and for a and 0 chains of hemoglobin in rabbit erythrocytes. ... 

Since all members of this family of RNA phages have homologous coat proteins, their subunits are expected to have the same three-dimensional structure. It remains to be seen if the MS2 fold is also present in any other unrelated viruses. The fold is so far unique for the MS2 subunit, but similar structures have been observed in other proteins such as the major histocompatibility antigen, HLA, which was discussed in Chapter 15. 

A majority of the work done in our laboratory has been conducted on E. coli and B. anthracis using MS2 and gamma-phage, respectively. Amplification results have been obtained from several other host bacteria and bacteriophages. Table 14.1 summarizes the materials studied. In all cases studied, no two bacteriophages contained the same proteins. 

The bacterial RNA viruses are all of quite small size, about 26 nm in size, and they are all icosahedral, with 180 copies of coat protein per virus particle. The complete nucleotide sequence of several RNA phages are known. In the RNA phage MS2, which infects Escherichia coli, the viral RNA is 3,569 nucleotides long. The virus RNA, although single stranded, has extensive regions of secondary and tertiary structure. The RNA strand in the virion has the plus (+) sense, acting directly as mRNA upon entry into the cell. 

Escherichia coli (strain JE24F+, derived from W3110, uninfected or infected with RNA-phage MS2 [2] recombinant overproducing strains [29,30] K-12 strains [30]) [2, 3, 28-31, 61, 62, 68, 70, 71]... 

Koizumi, Y. and Taya, M. (2002) Photocatalytic inactivation rate of phage MS2 in titanium dioxide suspensions containing various ionic species. Biotechnol. Lett. 24,459 162. 

Parts of the 2569-nucleotide sequence for the RNA of phage MS2 are shown in Fig. 29-17. The 5 end (upper left center) still bears the triphosphate group of the initiating GTE Following a number of hairpin loops there is a ribosome-protected region, which begins with the initiation codon GUG for the A protein... 

We have extensive test data where we challenged nano alirmina filters with various bacteria (E. coli, R. terrigina, B. globiggi, B. diminuta). We found that all such bacteria in the presence of a food source, can proliferate on the filter, and we therefore presume the filters are compatible with bacteria. We have also collected MS2 vims on NanoCeram and have been able to recover -90% of viable phage. The toxicity of raw nano alumina fiber towards E. coli, Staphylococcus aureus, B. subtilis, B. pumilis, and Candida albicans has been... 

Soon after T4 phage has infected E. coli, host protein synthesis is inhibited and the bacterium cannot be superinfected with RNA phage. This is reflected by the inability of ribosomes from T4 infected E. coli to translate MS2, f2, or R17 RNA in vitro or to bind to the initiation sites at the beginning of each cistron (Dube and Rudland, 1970 Steitz et al., 1970 Hsu and Weiss, 1969 Klem et al, 1970). If ribosomes from infected cells are washed in 2.0 M NH,C1 and then supplemented with initiation factors from normal ribosomes they are once more able to translate phage RNA. On the other hand, the wash fraction from infected ribosomes prevents, and even reverses, the formation of initiation complexes between normal ribosomes and MS2 RNA. These observations suggest that after T4 infection E. coli initiation factors are selectively... 

In some cases the rate of RNA folding may play a functional role and be exploited by natural systems. Such kinetic models were proposed for the control of translation by RNA secondary structure 28, 29). In these cases the translation is inhibited by a structure in the mRNA that sequesters the translation initiation region. Therefore the level of protein synthesis is very sensitive to the time of folding which actually regulates translation. For example, a long-distance interaction (LDI) was suggested to repress the synthesis of one of the proteins in phage MS2 (29). However,... 

Alkaline glutaraldehyde (2%) inhibits the transduction of P. aeruginosa PAO by bacteriophage FI 16, but is less effective in inactivating the phage [165]. Lower concentrations of the aldehyde (0.5%) reduce the titer of MS2-and K-coliphage by a factor of Iff within 20 minutes [166]. 

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