1887

Abstract

In the satellite phage P4, transcription starting from P is prevalently controlled via premature termination at several termination sites. We identified a spontaneous mutation, P4 (ppression of ermination), in the natural stop codon of P4 that, by elongating translation, suppresses transcription termination at the downstream site. Both the translational and the transcriptional profile of P4 differed from those of P4 wild-type. First of all, P4 did not express Orf151, but a higher molecular mass protein, compatible with the 303 codon open reading frame generated by the fusion of , and the intervening 138 nt. Moreover, after infection of , the mutant expressed a very low amount of the 1.3 and 1.7 kb transcripts originating at P and P promoters, respectively, and terminating at the intracistronic site, whereas correspondingly higher amounts of the 4.1 and 4.5 kb RNAs arising from the same promoters and covering the entire operon were detected. Thus the mutation converts a natural stop codon into a sense codon, suppresses a natural intracistronic termination site and leads to overexpression of the downstream and genes. This correlates with the inability of P4 to propagate in the plasmid state. By cloning different P4 DNA fragments, we mapped the transcription termination site within the 7633–7361 region between and gene . A potential stem–loop structure, resembling the structure of a Rho-independent termination site, was predicted by mfold sequence analysis at 7414–7385.

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2007-03-01
2024-03-28
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References

  1. Alano P., Dehò G., Sironi G., Zangrossi S. 1986; Regulation of the plasmid state of the genetic element P4. Mol Gen Genet 203:445–450 [CrossRef]
    [Google Scholar]
  2. Banerjee S., Chalissery J., Bandley I., Sen R. 2006; Rho-dependent transcription termination: more questions than answers. J Microbiol 44:11–22
    [Google Scholar]
  3. Borukhov S., Jookyung L., Laptenko O. 2005; Bacterial transcription elongation factors: new insights into molecular mechanism of action. Mol Microbiol 55:1315–1324
    [Google Scholar]
  4. Briani F., Zangrossi S., Ghisotti D., Dehò G. 1996; A Rho-dependent transcription termination site regulated by bacteriophage P4 RNA immunity factor. Virology 223:57–67 [CrossRef]
    [Google Scholar]
  5. Briani F., Ghisotti D., Dehò G. 2000; Antisense RNA-dependent transcription termination sites that modulate lysogenic development of satellite phage P4. Mol Microbiol 36:1124–1134 [CrossRef]
    [Google Scholar]
  6. Briani F., Dehò G., Forti F., Ghisotti D. 2001; The plasmid status of satellite bacteriophage P4. Plasmid 45:1–17 [CrossRef]
    [Google Scholar]
  7. Dehò G. 1983; Circular genetic map of satellite bacteriophage P4. Virology 126:267–278 [CrossRef]
    [Google Scholar]
  8. Dehò G., Ghisotti D. 2006; The satellite phage P4. In The Bacteriophages , 2nd edn. pp  391–408 Edited by Calendar R. New York: Oxford University Press;
    [Google Scholar]
  9. Dehò G., Ghisotti D., Alano P., Zangrossi S., Borrello M. G., Sironi G. 1984; Plasmid mode of propagation of the genetic element P4. J Mol Biol 178:191–207 [CrossRef]
    [Google Scholar]
  10. Dehò G., Zangrossi S., Ghisotti D., Sironi G. 1988; Alternative promoters in the development of bacteriophage plasmid. J Virol 62:1697–1704
    [Google Scholar]
  11. Dehò G., Zangrossi S., Sabbattini P., Sironi G., Ghisotti D. 1992; Bacteriophage P4 immunity controlled by small RNAs via transcription termination. Mol Microbiol 6:3415–3425 [CrossRef]
    [Google Scholar]
  12. Forti F., Polo S., Lane K. B., Six E. W., Sironi G., Dehò G., Ghisotti D. 1999; Translation of two nested genes in bacteriophage P4 controls immunity-specific transcription termination. J Bacteriol 181:5225–5233
    [Google Scholar]
  13. Forti F., Dragoni I., Briani F., Dehò G., Ghisotti D. 2002; Characterization of the small antisense CI RNA that regulates bacteriophage P4 immunity. J Mol Biol 315:541–549 [CrossRef]
    [Google Scholar]
  14. Ghisotti D., Finkel S., Halling C., Dehò G., Sironi G., Calendar R. 1990; Nonessential region of bacteriophage P4: DNA sequence, transcription, gene products, and functions. J Virol 64:24–36
    [Google Scholar]
  15. Ghisotti D., Chiaramonte R., Forti F., Zangrossi S., Sironi G., Dehò G. 1992; Genetic analysis of the immunity region of phage-plasmid P4. Mol Microbiol 6:3405–3413 [CrossRef]
    [Google Scholar]
  16. Henkin T. M., Yanofsky C. 2002; Regulation by transcription attenuation in bacteria: how RNA provides instructions for transcription termination/antitermination decisions. Bioessays 24:700–707 [CrossRef]
    [Google Scholar]
  17. Lesnik E. A., Sampath R., Levene H. B., Henderson T. J., McNeil J. A., Ecker D. J. 2001; Prediction of rho-independent transcriptional terminators in Escherichia coli . Nucleic Acids Res 29:3583–3594 [CrossRef]
    [Google Scholar]
  18. Lin C. S. 1984; Nucleotide sequence of the essential region of bacteriophage P4. Nucleic Acids Res 12:8667–8684 [CrossRef]
    [Google Scholar]
  19. Lindahl G., Sironi G., Bialy H., Calendar R. 1970; Bacteriophage lambda; abortive infection of bacteria lysogenic for phage P2. Proc Natl Acad Sci U S A 66:587–594 [CrossRef]
    [Google Scholar]
  20. Lindqvist B. H., Six E. W. 1971; Replication of bacteriophage P4 DNA in a nonlysogenic host. Virology 43:1–7 [CrossRef]
    [Google Scholar]
  21. Piazza F., Zappone M., Sana M., Briani F., Dehò G. 1996; Polynucleotide phosphorylase of Escherichia coli is required for the establishment of bacteriophage P4 immunity. J Bacteriol 178:5513–5521
    [Google Scholar]
  22. Piazzolla D., Calì S., Spoldi E., Forti F., Sala C., Magnoni F., Dehò G., Ghisotti D. 2006; Expression of phage P4 integrase is negatively regulated by both Int and Vis. J Gen Virol 87:2423–2431 [CrossRef]
    [Google Scholar]
  23. Reeve J. N. 1977; Bacteriophage infection of minicells: a general method for identification of ‘in vivo’ bacteriophage directed polypeptide biosynthesis. Mol Gen Genet 158:73–79 [CrossRef]
    [Google Scholar]
  24. Reeve J. 1979; Use of minicells for bacteriophage-directed polypeptide synthesis. Methods Enzymol 68:493–503
    [Google Scholar]
  25. Regonesi M. E., Briani F., Ghetta A., Zangrossi S., Ghisotti D., Tortora P., Dehò G. 2004; A mutation in polynucleotide phosphorylase from Escherichia coli impairing RNA binding and degradosome stability. Nucleic Acids Res 32:1006–1017 [CrossRef]
    [Google Scholar]
  26. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A 74:5463–5467 [CrossRef]
    [Google Scholar]
  27. Sauer B., Ow D., Ling L., Calendar R. 1981; Mutants of satellite bacteriophage P4 that are defective in the suppression of transcriptional polarity. J Mol Biol 145:29–46 [CrossRef]
    [Google Scholar]
  28. Sasaki I., Bertani G. 1965; Growth abnormalities in Hfr derivatives of Escherichia coli strain C. J Gen Microbiol 40:365–376 [CrossRef]
    [Google Scholar]
  29. Sunshine M. G., Thorn M., Gibbs W., Calendar R., Kelly B. 1971; P2 phage amber mutants: characterization by use of a polarity suppressor. Virology 46:691–702 [CrossRef]
    [Google Scholar]
  30. Terzano S., Christian R., Espinoza F. H., Calendar R., Dehò G., Ghisotti D. 1994; A new gene of bacteriophage P4 that controls DNA replication. J Bacteriol 176:6059–6065
    [Google Scholar]
  31. Tocchetti A., Serina S., Oliva I., Dehò G., Ghisotti D. 2001; Cnr interferes with dimerization of the replication protein alpha in phage-plasmid P4. Nucleic Acids Res 29:536–544 [CrossRef]
    [Google Scholar]
  32. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119 [CrossRef]
    [Google Scholar]
  33. Ziegelin G., Calendar R., Ghisotti D., Terzano S., Lanka E. 1997; Cnr protein, the negative regulator of bacteriophage P4 replication, stimulates specific DNA binding of its initiator protein alpha. J Bacteriol 179:2817–2822
    [Google Scholar]
  34. Zuker M. 2003; Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res 31:3406–3415 [CrossRef]
    [Google Scholar]
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