메뉴 건너뛰기




Volumn 284, Issue 5414, 1999, Pages 611-615

Mechanism of intrinsic transcription termination and antitermination

Author keywords

[No Author keywords available]

Indexed keywords

DNA HYBRID; RNA; RNA POLYMERASE;

EID: 0033597435     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.284.5414.611     Document Type: Article
Times cited : (282)

References (52)
  • 16
    • 0029029468 scopus 로고
    • D. Wang et al., Cell 81, 341 (1995).
    • (1995) Cell , vol.81 , pp. 341
    • Wang, D.1
  • 26
    • 0345394594 scopus 로고    scopus 로고
    • note
    • 2, and poly(dA-dT) (20 μg/ml)], resuspended in TB2 plus 100 nM NusA (50), divided into samples containing 10 nM complex, and supplemented with 150 μM oligo where added. After addition of 100 μM ATP and GTP, 10 μM CTP, and 50 μM UTP, samples were removed at indicated times during incubation at 37°C, divided into supernatant and pellet by magnetic partitioning, and diluted with stop buffer [final concentration 500 mM tris-HCl (pH 7.9), 10 mM EDTA, tRNA (50 μg/ml)]. After extraction with phenol/chloroform/isoamyl alcohol and ethanol precipitation, samples were resolved on a 7% polyacrylamide gel, imaged by Molecular Dynamics Phosphorimager, and quantified by ImageQuant software.
  • 27
    • 0345394593 scopus 로고    scopus 로고
    • note
    • I. Artsimovitch and R. Landick (51) have shown release of RNA by an oligo at an attenuator-related pause site.
  • 28
    • 0345394590 scopus 로고    scopus 로고
    • note
    • 2), followed by advance to +76 by incubation for 2 min at 37°C in TB4 plus 150 nM NusA and 25μM CTP, GTP, and ATP; most complexes advance to +77. The release of transcripts by oligos t2, t3, and t4 was measured during incubation of B nM complexes in TB4 plus 150 nM NusA for 5 min at 37°C with 100 μM oligo followed by the addition of 250 nM ATP, GTP, CTP, and UTP (which allowed very slow synthesis) for 15 min. For release with oligo t5, washed +76 complexes were spread downstream of +76 by incubation with 100 nM ATP and GTP and 2.5 μM UTP for 1.5 min, followed by four washes and incubation for 5 min with 200 μM oligo. Some complexes incubated without NTPs but with oligo were found to be arrested, failing to elongate upon further incubation with NTP (10, 14, 17), so release efficiency of the unarrested fraction also was calculated. Supernatant and pellet fractions were determined as before, and the percent transcript release was calculated.
  • 34
    • 0344532286 scopus 로고    scopus 로고
    • note
    • 2), advanced to +91 in TB2 plus 150 nM NusA, 15 μM GTP, 15 μM ATP, and 15 μM UTP (or six times these concentrations of NTPs for RpoB8), and spread downstream of +91 by incubation for 6 min at 37°C in TB2 with 750 nM ATP and GTP, 250 nM UTP, and 40 nM CTP (six times these concentrations of NTPs for RpoB8). After five washes in TB6, complexes were resuspended in TB2 plus 150 nM NusA, incubated 0.5 min at 37°C, supplemented with 100 μM oligo t8 (complementary to the transcript from +86 to +105) or t8a (as t8, but mismatched at +99 and +101), and sampled at 4 and 16 min. There was no detectable release by t8a.
  • 35
    • 0000483406 scopus 로고
    • S. L. McKnight and K. R. Yamamoto, Eds. (Cold Spring Harbor Laboratory Press, Plainview, NY)
    • J. W. Roberts, in Transcriptional Regulation, Book 1, S. L. McKnight and K. R. Yamamoto, Eds. (Cold Spring Harbor Laboratory Press, Plainview, NY, 1992), p. 389.
    • (1992) Transcriptional Regulation, Book 1 , pp. 389
    • Roberts, J.W.1
  • 38
    • 0345394591 scopus 로고    scopus 로고
    • This experiment is described in supplementary material available at
    • This experiment is described in supplementary material available at www.sciencemag.org/feature/data/ 987284.shl.
  • 41
    • 0344963183 scopus 로고    scopus 로고
    • note
    • 2), suspended in TB5 plus 150 nM NusA, incubated with oligo at 200 μM for 16 min at 37°C, and assayed for release.
  • 42
    • 0031558773 scopus 로고    scopus 로고
    • W. A. Rees, S. E. Weitzel, A. Das, P. H. von Hippel [J. Mol. Biol. 273, 797 (1997)] suggest that the λ gene N antiterminator also stabilizes transcription complexes, on the basis of reaction kinetics at nonrelease sites.
    • (1997) J. Mol. Biol. , vol.273 , pp. 797
    • Rees, W.A.1    Weitzel, S.E.2    Das, A.3    Von Hippel, P.H.4
  • 45
    • 0344963181 scopus 로고    scopus 로고
    • note
    • 82; bound and released RNAs were measured during incubation. An identical result was obtained with a t500 derivative lacking the U-rich segment.
  • 46
    • 0344100833 scopus 로고    scopus 로고
    • note
    • A related forward translocation model was proposed by Chamberlin (12).
  • 52
    • 0344532281 scopus 로고    scopus 로고
    • note
    • Supported by NIH grant GM 21941. We thank R. Weisberg and L. Rothman-Denes for reading the manuscript, members of the laboratory, especially M. Marr, for essential advice and discussion, and D. Jun-Jin for a gift of mutationally altered RNA polymerase.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.