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PMID: 7521531 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Control of transcription processivity in phage lambda: Nus factors strengthen the termination-resistant state of RNA polymerase induced by N antiterminator.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 91 ·No. 18 ·1994-08-30 ·Pages 8660-4

DeVito J, Das A

Abstract

During transcription of phage lambda early operons, the N gene product alters host RNA polymerase (RNAP) so that transcription proceeds through multiple stop signals. Here, we reproduce the essence of N activity with purified components in synthetic transcription units that contain lambda pL promoter and the N-recognition site, nutL, followed by a variety of intrinsic terminators. We show that three host factors (NusA, NusE, and NusG) are essential for N to allow appreciable transcription through multiple terminators and that this persistent antitermination is stimulated by a fourth factor, NusB. Remarkably, in the absence of all four factors, N suppresses various terminators placed near the nut site. This basal antitermination activity of N is enhanced by NusA and is diminished by high salt and temperature. We postulate that N interacts with RNAP directly, inducing the termination-resistant state. While NusA facilitates this interaction, the other factors strengthen it sufficiently over time and distance so that RNAP bypasses multiple terminators. The dispensability of NusB for persistent antitermination in vitro, but not in vivo, raises the possibility that NusB performs two functions: it increases the stability of N antitermination complex and also counteracts an inhibitory factor in the cell.

MeSH Terms
Bacterial Proteins/physiology Bacteriophage lambda/genetics DNA-Directed RNA Polymerases/metabolism Escherichia coli Proteins In Vitro Techniques Peptide Elongation Factors/physiology RNA, Bacterial/biosynthesis Ribosomal Proteins/physiology Terminator Regions, Genetic Transcription Factors/physiology Transcription, Genetic Transcriptional Elongation Factors Viral Regulatory and Accessory Proteins/physiology
Chemicals
Bacterial Proteins Escherichia coli Proteins N protein, Bacteriophage lambda NusB protein, E coli NusG protein, E coli Peptide Elongation Factors RNA, Bacterial Ribosomal Proteins Transcription Factors Transcriptional Elongation Factors Viral Regulatory and Accessory Proteins nusA protein, E coli ribosomal protein S10 DNA-Directed RNA Polymerases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
DeVito J
Department of Microbiology, University of Connecticut Health Center, Farmington 06030.
Das A
References (46)
46 references, click to expand
  1. Replacement of potassium chloride by potassium glutamate dramatically enhances protein-DNA interactions in vitro.
    Biochemistry. 1987 Apr 21;26(8):2095-101 PMID: 2887198
  2. Interaction of the sigma factor and the nusA gene protein of E. coli with RNA polymerase in the initiation-termination cycle of transcription.
    Cell. 1981 May;24(2):421-8 PMID: 6263495
  3. An elongation control particle containing the N gene transcriptional antitermination protein of bacteriophage lambda.
    Cell. 1987 Nov 20;51(4):631-41 PMID: 2445491
  4. Functional elements of DNA upstream from the integrase operon that are conserved in bacteriophages 434 and lambda.
    Gene. 1987;61(2):135-44 PMID: 2965063
  5. NusA protein is necessary and sufficient in vitro for phage lambda N gene product to suppress a rho-independent terminator placed downstream of nutL.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2494-8 PMID: 2965813
  6. Effects of rifampicin resistant rpoB mutations on antitermination and interaction with nusA in Escherichia coli.
    J Mol Biol. 1988 Nov 20;204(2):247-61 PMID: 2464690
  7. Sequence-specific recognition of RNA hairpins by bacteriophage antiterminators requires a conserved arginine-rich motif.
    Cell. 1989 Oct 6;59(1):207-18 PMID: 2477156
  8. Transcriptional antitermination in the bgl operon of E. coli is modulated by a specific RNA binding protein.
    Cell. 1990 Sep 21;62(6):1153-63 PMID: 1698125
  9. Transcription-dependent competition for a host factor: the function and optimal sequence of the phage lambda boxA transcription antitermination signal.
    Genes Dev. 1990 Dec;4(12A):2210-22 PMID: 2148536
  10. Action of an RNA site at a distance: role of the nut genetic signal in transcription antitermination by phage-lambda N gene product.
    New Biol. 1990 Nov;2(11):975-91 PMID: 2151659
  11. Assembly of transcription elongation complexes containing the N protein of phage lambda and the Escherichia coli elongation factors NusA, NusB, NusG, and S10.
    Genes Dev. 1991 Aug;5(8):1504-12 PMID: 1831176
  12. RNA polymerase: regulation of transcript elongation and termination.
    FASEB J. 1991 Oct;5(13):2833-42 PMID: 1916107
  13. The nut site of bacteriophage lambda is made of RNA and is bound by transcription antitermination factors on the surface of RNA polymerase.
    Genes Dev. 1991 Nov;5(11):2141-51 PMID: 1834523
  14. The Psu protein of bacteriophage P4 is an antitermination factor for rho-dependent transcription termination.
    J Bacteriol. 1991 Nov;173(21):6722-31 PMID: 1938879
  15. Specificity of antitermination mechanisms. Suppression of the terminator cluster T1-T2 of Escherichia coli ribosomal RNA operon, rrnB, by phage lambda antiterminators.
    J Mol Biol. 1991 Nov 5;222(1):59-66 PMID: 1719220
  16. Direct interaction between two Escherichia coli transcription antitermination factors, NusB and ribosomal protein S10.
    J Mol Biol. 1992 Jan 5;223(1):55-66 PMID: 1731086
  17. Effect of Escherichia coli nusG function on lambda N-mediated transcription antitermination.
    J Bacteriol. 1992 Feb;174(4):1339-44 PMID: 1531224
  18. Requirement for E. coli NusG protein in factor-dependent transcription termination.
    Cell. 1992 Mar 6;68(5):989-94 PMID: 1547498
  19. Parameters affecting transcription termination by Escherichia coli RNA polymerase. I. Analysis of 13 rho-independent terminators.
    J Mol Biol. 1992 Mar 5;224(1):31-51 PMID: 1372365
  20. Simultaneous gain and loss of functions caused by a single amino acid substitution in the beta subunit of Escherichia coli RNA polymerase: suppression of nusA and rho mutations and conditional lethality.
    Genetics. 1992 Mar;130(3):411-28 PMID: 1551568
  21. The phage lambda gene Q transcription antiterminator binds DNA in the late gene promoter as it modifies RNA polymerase.
    Cell. 1992 Jun 26;69(7):1181-9 PMID: 1535556
  22. Host factor requirements for processive antitermination of transcription and suppression of pausing by the N protein of bacteriophage lambda.
    J Biol Chem. 1992 Sep 25;267(27):19418-26 PMID: 1388170
  23. How the phage lambda N gene product suppresses transcription termination: communication of RNA polymerase with regulatory proteins mediated by signals in nascent RNA.
    J Bacteriol. 1992 Nov;174(21):6711-6 PMID: 1400223
  24. Elongation factor NusG interacts with termination factor rho to regulate termination and antitermination of transcription.
    Genes Dev. 1993 Jan;7(1):161-72 PMID: 8422985
  25. Recognition of boxA antiterminator RNA by the E. coli antitermination factors NusB and ribosomal protein S10.
    Cell. 1993 Jan 29;72(2):261-8 PMID: 7678781
  26. Ribosomal RNA antitermination in vitro: requirement for Nus factors and one or more unidentified cellular components.
    Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):970-4 PMID: 8430111
  27. Transcriptional antitermination.
    Nature. 1993 Jul 29;364(6436):401-6 PMID: 8332211
  28. Control of transcription termination by RNA-binding proteins.
    Annu Rev Biochem. 1993;62:893-930 PMID: 8352604
  29. The nusA gene protein of Escherichia coli. Its identification and a demonstration that it interacts with the gene N transcription anti-termination protein of bacteriophage lambda.
    J Mol Biol. 1981 Mar 25;147(1):11-23 PMID: 6455533
  30. Transcription antitermination in vitro by lambda N gene product: requirement for a phage nut site and the products of host nusA, nusB, and nusE genes.
    Cell. 1984 Aug;38(1):165-73 PMID: 6088061
  31. nusB: a protein factor necessary for transcription antitermination in vitro by phage lambda N gene product.
    Proc Natl Acad Sci U S A. 1984 Oct;81(20):6305-9 PMID: 6093096
  32. Interactions of bacteriophage and host macromolecules in the growth of bacteriophage lambda.
    Microbiol Rev. 1984 Dec;48(4):299-325 PMID: 6240590
  33. Evidence that ribosomal protein S10 itself is a cellular component necessary for transcription antitermination by phage lambda N protein.
    Proc Natl Acad Sci U S A. 1985 Jun;82(12):4070-4 PMID: 2987961
  34. Phage lambda gene Q antiterminator recognizes RNA polymerase near the promoter and accelerates it through a pause site.
    Cell. 1985 Aug;42(1):259-69 PMID: 2990726
  35. Bacteriophage lambda N-dependent transcription antitermination. Competition for an RNA site may regulate antitermination.
    J Mol Biol. 1994 Feb 11;236(1):217-28 PMID: 8107107
  36. Termination factor for RNA synthesis.
    Nature. 1969 Dec 20;224(5225):1168-74 PMID: 4902144
  37. Bacterial mutants in which the gene N function of bacteriophage lambda is blocked have an altered RNA polymerase.
    Proc Natl Acad Sci U S A. 1971 Dec;68(12):2977-81 PMID: 4943550
  38. Relationship between the N function of bacteriophage lambda and host RNA polymerase.
    J Mol Biol. 1972 Mar 28;65(2):259-72 PMID: 5040359
  39. Gene N regulator function of phage lambda immun21: evidence that a site of N action differs from a site of N recognition.
    J Mol Biol. 1973 Dec 25;81(4):505-16 PMID: 4778807
  40. Genetic characterization of a bacterial locus involved in the activity of the N function of phage lambda.
    Virology. 1974 Mar;58(1):141-8 PMID: 4595374
  41. Cooperative effects of bacterial mutations affecting lambda N gene expression. I. Isolation and characterization of a nusB mutant.
    Virology. 1976 Aug;73(1):119-27 PMID: 785802
  42. The relationship between function and DNA sequence in an intercistronic regulatory region in phage lambda.
    Nature. 1978 Mar 30;272(5652):414-23 PMID: 634366
  43. Coliphage lambdanutL-: a unique class of mutants defective in the site of gene N product utilization for antitermination of leftward transcription.
    J Mol Biol. 1978 Sep 5;124(1):195-221 PMID: 361968
  44. Transcription antitermination by bacteriophage lambda N gene product.
    J Mol Biol. 1980 Jun 15;140(1):57-75 PMID: 6447798
  45. Evidence that ribosomal protein S10 participates in control of transcription termination.
    Proc Natl Acad Sci U S A. 1981 Feb;78(2):1115-8 PMID: 6453343
  46. An antitermination protein engages the elongating transcription apparatus at a promoter-proximal recognition site.
    Cell. 1987 Sep 11;50(6):885-99 PMID: 3040263
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.91.18.8660
Published
1994-08-30
Pages
8660-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC44666
Subset
IM
Grants
NIGMS NIH HHS · GM28946 · United States
External Links
PubMed source DOI: 10.1073/pnas.91.18.8660 PMC Full Text View journal details
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