Protein p4 represses phage phi 29 A2c promoter by interacting with the alpha subunit of Bacillus subtilis RNA polymerase (original) (raw)

1996, Proceedings of the National Academy of Sciences

Abstract

Regulatory protein p4 from Bacillus subtilis phage phi 29 represses the strong viral A2c promoter (PA2c) by preventing promoter clearance; it allows RNA polymerase to bind to the promoter and form an initiated complex, but the elongation step is not reached. Protein p4 binds at PA2c immediately upstream from RNA polymerase; repression involves a contact between both proteins that holds the RNA polymerase at the promoter. This contact is held mainly through p4 residue Arg120, which is also required for activation of the phi 29 late A3 promoter. We have investigated which region of RNA polymerase contacts protein p4 at PA2c. Promoter repression was impaired when a reconstituted RNA polymerase lacking the 15 C-terminal residues of the alpha subunit C-terminal domain was used; this polymerase was otherwise competent for transcription. Binding cooperativity assays indicated that protein p4 cannot interact with this mutant RNA polymerase at PA2c. Protein p4 could form a complex at PA2c wi...

Loading...

Loading Preview

Sorry, preview is currently unavailable. You can download the paper by clicking the button above.

References (39)

  1. Yura, T. & Ishihama, A. (1979) Annu. Rev. Genet. 13, 59-97.
  2. Jin, R., Sharif, K. A. & Krakow, J. S. (1995) J. Biol. Chem. 270, 19213-19216.
  3. Baldus J. M., Buckner, C. & Moran, C. P. (1995) Mol. Microbiol. 17, 281-290.
  4. Li, M., Moyle, H. & Susskind, M. M. (1994) Science 263, 75-77.
  5. Makino, K., Amemura, M., Kim, S.-K., Nakata, A. & Shinegawa, H. (1993) Genes Dev. 7, 149-170.
  6. Kumar, A., Grimes, B., Fujita, N., Makino, K., Malloch, R. A., Hayward, R. & Ishihama, A. (1994) J. Mo. Bio. 235, 405-413.
  7. Kuldell, N. & Hochschild, A. (1994) J. Bacteriol. 176, 2991-2998.
  8. Ishihama, A. (1993) J. Bacteriol. 175, 2483-2489.
  9. Ebright, R. E. & Busby, S. (1995) Curr. Opin. Genet. Dev. 5, 197-203.
  10. Blatter, E. E., Ross, W., Tang, H., Gourse, R. L. & Ebright, R. H. (1994) Cell 78, 889-896.
  11. Negishi, T., Fujita, N. & Ishihama, A. (1995) J. Mo. Biol. 248, 723-728.
  12. Hayward, R., Igarashi, K. & Ishihama, A. (1991) J. Mol. Bio. 221, 23-29.
  13. Igarashi, K., Fujita, N. & Ishihama, A. (1991) J. Mol. Bio. 218, 1-6.
  14. Kimura, M., Fujita, N. & Ishihama, A. (1994) J. Mo. Biol. 242, 107-115.
  15. Tang, H., Severinov, K., Goldfarb, A., Fenyo, D., Chait, B. & Ebright, R. E. (1994) Genes Dev. 8, 3058-3067.
  16. Mencia, M., Monsalve, M., Rojo, F. & Salas, M. (1996) Proc. Natl. Acad. Sci. USA 93, in press.
  17. Ross, W., Gosink, K. K., Salomon, J., Igarashi, K., Zou, C., Ishihama, A., Severinov, K. & Gourse, R. L. (1993) Science 262, 1407-1413.
  18. Rao, L., Ross, W., Appleman, J. A., Gaal, T., Leirmo, S., Schlax, P. J., Record, M. T. & Gourse, R. L. (1994) J. Mo. Bio. 235, 1421-1435.
  19. Fredrick, K., Caramori, T., Chen, Y.-F., Galizzi, A. & Helmann, J. (1995) Proc. Natl. Acad. Sci. USA 92, 2582-2586.
  20. Choy, H. E., Park, S. W., Aki, T., Parrack, P., Fujita, N., Ishi- hama, A. & Adhya, S. (1995) EMBO J. 14, 4523-4529.
  21. Monsalve, M., Mencia, M., Rojo, F. & Salas, M. (1996) EMBO J. 15, 383-391.
  22. Sogo, J. M., Inciarte, M. R., Corral, J., Vifiuela, E. & Salas, M. (1979) J. Mo. Biol. 127, 411-436.
  23. Mencia, M., Salas, M. & Rojo, F. (1993) J. Mol. Biol. 233, 695-704.
  24. Chang, B. Y. & Doi, R. H. (1990) J. Bacteriol. 172, 3257-3263.
  25. Monsalve, M., Mencia, M., Rojo, F. & Salas, M. (1995) Virology 207, 23-31.
  26. Lane, D., Prentki, P. & Chandler, M. (1992) Microbiol. Rev. 56, 509-528.
  27. Mencia, M., Monsalve, M., Salas, M. & Rojo, F. (1996) Mo. Microbiol. 20, 273-282.
  28. Jeon, Y. H., Negishi, T., Shirakawa, M., Yamazaki, T., Fujita, N., Ishihama, A. & Kyogoku, Y. (1995) Science 270, 1495-1497.
  29. Sander, C. & Schneider, R. (1991) Proteins 9, 56-68.
  30. Gaal, T., Ross, W., Blatter, E. E., Tang, H., Jia, X., Krishnan, V. V., Assa-Munt, N., Ebright, R. H. & Gourse, R. L. (1996) Genes Dev. 10, 16-26.
  31. Zhou. Y., Pendergrast, P. S., Bell, A., Williams, R., Busby, S. & Ebright, R. H. (1994) EMBO J. 13, 4549-4557.
  32. Barthelemy, I. & Salas, M. (1989) J. Mol. Biol. 208, 225-232.
  33. Rojo, F. & Salas, M. (1991) EMBO J. 10, 3429-3438.
  34. Nuez, B., Rojo, F. & Salas, M. (1992) Proc. Natl. Acad. Sci. USA 89, 11401-11405.
  35. Gaston, K., Bell, A., Kolb, A., Buc, H. & Busby, S. (1990) Cell 62, 733-743.
  36. Ushida, C. & Aiba, H. (1990) Nucleic Acids Res. 18, 6325-6330.
  37. Wing, H. J., Williams, S. M. & Busby, S. J. W. (1995) J. Bacteriol. 177, 6704-6710.
  38. Ellinger, T., Behnke, D., Bujard, H. & Gralla, J. (1994) J. Mol. Biol. 239, 455-465.
  39. Proc. Natl. Acad. Sci. USA 93 (1996)