RNA polymerase β' subunit: A target of DNA binding-independent activation

Science

Volumn 275, Issue 5306, 1997, Pages 1655-1657

  Miller, Alita ^a,c   Wood, Douglas ^a   Ebright, Richard H ^b   Rothman Denes, Lucia B ^a  

^a UNIVERSITY OF CHICAGO   (United States)

^b RUTGERS UNIVERSITY   (United States)

^c INSTITUT DE GÉNÉTIQUE ET DE BIOLOGIE MOLÉCULAIRE ET CELLULAIRE   (France)

Author keywords

[No Author keywords available]

Indexed keywords

DNA BINDING PROTEIN; RNA POLYMERASE;

ARTICLE; DNA BINDING; ESCHERICHIA COLI; NONHUMAN; PRIORITY JOURNAL; PROMOTER REGION; PROTEIN DNA INTERACTION; TRANSCRIPTION INITIATION;

ESCHERICHIA COLI; EUKARYOTA; PROKARYOTA;

EID: 0030896970     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.275.5306.1655     Document Type: Article

References (43)

1. N.-Y. Cho, M. Choi, L. B. Rothman-Denes, J. Mol. Biol. 246, 461 (1995).
2. G. J. Lindberg, S. C. Kowalczykowski, J. K. Rist, A. Sugino, L. B. Rothman-Denes, J. Biol. Chem. 264, 12700 (1989).
3. M. Choi, A. Miller, N.-Y. Cho, L. B. Rothman-Denes, ibid. 270, 22541 (1995).
4. A. Miller, D. Wood, M. Choi, L. B. Rothman-Denes, unpublished data.
5. 128 was substituted with Ala; in Δ264-265;S260A, residues 264 and 265 were deleted and Ser (S) at position 260 was substituted with Ala; and in K264A;K265A, residues Lys (K) at positions 264 and 265 were substituted with Ala.
6. 3H]thymidine incorporation), before or after induction with isopropyl-thio-β-D-galactoside (IPTG) (1, 3). The ability of N4SSB and its derivatives to activate transcription in vitro was measured in run-off transcription experiments with an 800-base pair (bp) DNA fragment containing the N4 late promoter R (which uses three start sites), RNAP, and increasing amounts of N4SSB (7). The ssDNA binding was assayed in electrophoretic mobility shift experiments (3). N4SSB and its derivatives were purified as in (7).
7. A. Miller, X. Dai, M. Choi, A. Glucksmann-Kuis, L. B. Rothman-Denes, Methods Enzymol. 174, 9 (1996).
8. 2-terminus of the a subunit.
9. M. Kashlev, E. Martin, K. Severinov, V. Nikiforov, A. Goldfarb, Gene 130, 9 (1993); H. Tang, K. Severinov, A. Goldfarb, R. H. Ebright, Proc. Natl. Acad. Sci. U.S.A. 92, 4902 (1995).
10. M. Kashlev, E. Martin, K. Severinov, V. Nikiforov, A. Goldfarb, Gene 130, 9 (1993); H. Tang, K. Severinov, A. Goldfarb, R. H. Ebright, Proc. Natl. Acad. Sci. U.S.A. 92, 4902 (1995).
11. R. Zivin, W. Zehring, L. B. Rothman-Denes, J. Mol. Biol. 152, 335 (1981).
12. L. A. Isaksson, S. E. Skold, J. Skjoldebrand, R. Takata, Mol. Gen. Genet. 156, 233 (1977).
13. G. E. Christie et al., J. Bacteriol. 178, 6991 (1996).
14. 620) of 0.4 and infected with wild-type N4 phage at a multiplicity of infection of 10. After 40 min, total RNA was harvested and tested for the presence of the N4 late promoter R and rrnB P1 transcripts by primer extension (1, 19).
15. C. A. Josaitis, T. Gaal, R. L. Gourse, Proc. Natl. Acad. Sci. U.S.A. 92, 1117 (1995); S. Borukhov, V. Sagitov, C. A. Josaitis, R. L. Gourse, A. Goldfarb, J. Biol. Chem. 268, 23477 (1993).
16. C. A. Josaitis, T. Gaal, R. L. Gourse, Proc. Natl. Acad. Sci. U.S.A. 92, 1117 (1995); S. Borukhov, V. Sagitov, C. A. Josaitis, R. L. Gourse, A. Goldfarb, J. Biol. Chem. 268, 23477 (1993).
17. Y. Chen, Y. W. Ebright, R. H. Ebright, Science 265, 90 (1994); Y. Ebright, Y. Chen, Y. Kim, R. H. Ebright, Bioconjugate Chem. 7, 380 (1996).
18. Y. Chen, Y. W. Ebright, R. H. Ebright, Science 265, 90 (1994); Y. Ebright, Y. Chen, Y. Kim, R. H. Ebright, Bioconjugate Chem. 7, 380 (1996).
19. W. Niu, Y. Kim, G. Tau, T. Heyduk, R. H. Ebright, Cell 87, 1123 (1996).
20. 125I-AET), photo cross-linking, cleavage after photo cross-linking, and radiolabel transfer were performed as in (15, 16). Reaction mixtures for photo cross-linking contained 250 nM N4SSB derivative and 125 nM RNAP (Epicentre Biotechnologies, Madison, WI). Control experiments with wild-type N4SSB with the cross-linking agent incorporated at C84 and C86 yielded no N4SSB-RNAP cross-linking.
21. 54-RNAP activate transcription in the absence of DNA binding; however, their function at physiological concentrations requires DNA binding [E. Huala and S. M. Ausubel, J. Bacteriol. 171, 3354 (1989); D. K. Berger, S. Naberhaus, S. Kustu, Proc. Natl. Acad. Sci. U.S.A. 91, 103 (1994); A. North and S. Kustu, J. Mol. Biol., in press].
22. 54-RNAP activate transcription in the absence of DNA binding; however, their function at physiological concentrations requires DNA binding [E. Huala and S. M. Ausubel, J. Bacteriol. 171, 3354 (1989); D. K. Berger, S. Naberhaus, S. Kustu, Proc. Natl. Acad. Sci. U.S.A. 91, 103 (1994); A. North and S. Kustu, J. Mol. Biol., in press].
23. 54-RNAP activate transcription in the absence of DNA binding; however, their function at physiological concentrations requires DNA binding [E. Huala and S. M. Ausubel, J. Bacteriol. 171, 3354 (1989); D. K. Berger, S. Naberhaus, S. Kustu, Proc. Natl. Acad. Sci. U.S.A. 91, 103 (1994); A. North and S. Kustu, J. Mol. Biol., in press].
24. A. Barberis et al., Cell 81, 359 (1995); S. Farrell et al., Genes Dev. 10, 2359 (1996); S. L. Dove, J. K. Joung, A. Hochschild, unpublished data.
25. A. Barberis et al., Cell 81, 359 (1995); S. Farrell et al., Genes Dev. 10, 2359 (1996); S. L. Dove, J. K. Joung, A. Hochschild, unpublished data.
26. N.-Y. Cho, A. Miller, L. B. Rothman-Denes, unpublished data.
27. S. Busby and R. H. Ebright, Cell 79, 743 (1994).
28. M. Li, H. Moyle, M. M. Susskind, Science 263, 75 (1994); N. Kudell and A. Hochschild, J. Bacteriol. 176, 2991 (1994).
29. M. Li, H. Moyle, M. M. Susskind, Science 263, 75 (1994); N. Kudell and A. Hochschild, J. Bacteriol. 176, 2991 (1994).
30. R. Jokerst, J. Weeks, W. Zehring, A. Greenleaf, Mol. Gen. Genet. 215, 266 (1989).
31. J. L. Corden and C. J. Ingles, in Transcriptional Regulation, S. L. McKnight and K. R. Yamamoto, Eds. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1992), pp. 81-108; A. Emili and C. J. Ingles, Curr. Opin. Genet. Dev. 5, 204 (1995).
32. J. L. Corden and C. J. Ingles, in Transcriptional Regulation, S. L. McKnight and K. R. Yamamoto, Eds. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1992), pp. 81-108; A. Emili and C. J. Ingles, Curr. Opin. Genet. Dev. 5, 204 (1995).
33. G. D. Meredith et al., J. Mol. Biol. 258, 413 (1996); K. K. Leuther, D. A. Bushnell, R. D. Kornberg, Cell 85, 773 (1996).
34. G. D. Meredith et al., J. Mol. Biol. 258, 413 (1996); K. K. Leuther, D. A. Bushnell, R. D. Kornberg, Cell 85, 773 (1996).
35. M. L. Nonet and R. A. Young, Genetics 123, 715 (1989).
36. C. Martin, S. Okamura, R. Young, Mol. Cell. Biol. 10, 1908 (1990).
37. H. Xiao, J. D. Friesen, J. T. Lis, ibid. 14, 7507 (1994).
38. C. Scale et al., Nature 374, 491 (1990); H. P. Gerber et al., ibid. 373, 660 (1995).
39. C. Scale et al., Nature 374, 491 (1990); H. P. Gerber et al., ibid. 373, 660 (1995).
40. T. O'Brien, S. Hardin, A. Greenleaf, J. T. Lis, ibid. 370, 75 (1994); A. Akhtar, G. Paye, D. J. Bentley, EMBO J. 15, 4654 (1996).
41. T. O'Brien, S. Hardin, A. Greenleaf, J. T. Lis, ibid. 370, 75 (1994); A. Akhtar, G. Paye, D. J. Bentley, EMBO J. 15, 4654 (1996).
42. C. J. Saris, J. van Eenbergen, B. G. Jenks, H. P. Bioemers, Anal. Biochem. 132, 54 (1983).
43. We thank R. Calendar, R. Landick, and W. Ross for strains and protein samples, G. Greene for facilities, and D. Gottschling, J. Greenblatt, G. Gussin, A. Hochschild, R. Landick, J. Lis, and R. Young for discussions. Supported by NIH grants GM35170 and GM54431 (to L.B.R.-D.) and GM41376 (to R.H.E.). A.M. was partially supported by USPHS grant 5T32 GM 07183.