Evidence that ribosomal protein S10 itself is a cellular component necessary for transcription antitermination by phage lambda N protein.

Das, Asis; Ghosh, Balaram; Barik, Sailen; Wolska, Krystyna I.

doi:10.1073/pnas.82.12.4070

Cited by 29 publications

(15 citation statements)

References 34 publications

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“…1). A set of bacterial proteins called Nus, which takes part in cellular transcriptional and translational processes, interacts with N and RNA Pol during N-mediated transcription antitermination (10,12,40,61). BoxA and BoxB are elements of the RNA Nut sites (24).…”

Section: Fig 2 Looping Of the Operators Ol And Or During CI Bindingmentioning

confidence: 99%

A New Look at Bacteriophage λ Genetic Networks

2007

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Section: Fig 2 Looping Of the Operators Ol And Or During CI Bindingmentioning

confidence: 99%

A New Look at Bacteriophage λ Genetic Networks

2007

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“…Antitermination by N is influenced by multiple protein factors of Escherichia coli (7,(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30). Three of these factors, NusA, NusB, and NusE (the S10 ribosomal protein), were discovered by the isolation of host mutants that are defective in N antitermination and, hence, fail to support A development (20)(21)(22).…”

Section: -18)mentioning

confidence: 99%

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

DeVito

Das²

1994

Proc. Natl. Acad. Sci. U.S.A.

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During transcription of phage A 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 A pL promoter and the N-recognition site, nutL, followed by a variety ofintrinsic 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 antitenation 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 terminationresistant 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.The RNA polymerase (RNAP) elongation complex is the target of two kinds of regulatory factors that modulate the elongation-termination decision in transcription (1-3). (4,5). Others convert RNAP to the termination-resistant form. The archetype of this latter class of antiterminators is the N gene product of phage A (6-8). The N protein is essential for transcription of the early genes of phage A, which are organized in two large operons (8). RNAP transcribing each operon encounters both factor-dependent and factor-independent terminators that are suppressed by N (9). The suppression of multiple terminators in each operon requires a specific site called nut, located between the promoter and the first terminator (10-12). The nut site acts in the form of RNA (13-15). During transcription, N is thought to bind to a small hairpin component of nut RNA, known as boxB, so that N can capture the RNAP elongation complex and transform RNAP into the termination-resistant form (7, 13-18).Antitermination by N is influenced by multiple protein factors of Escherichia coli (7,(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30). Three of these factors, NusA, NusB, and NusE (the S10 ribosomal protein), were discovered by the isolation of host mutants that are defective in N antitermination and, hence, fail to support A development (20-22). The fourth factor, NusG, emerged from the isolation ofextragenic suppressors, which permit A growth in a nusA mutant host (28). Further genetic studies indicated that these factors interact with each other, N, and RNAP (19). Indeed, N and all four host factors form a stable transcription complex in vitro (16,17,29). Although no evidence for a direct interaction between N and RNAP has been reported, N does bin...

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“…Ribosomal protein S10 is an intrinsic part of the antiterruination reaction, because mutations in S10 prevent antitermination in vivo (Friedman et al 1981) and in vitro (Das et al 1985;Horwitz et al 1987). The role of S10 in antitermination by N may be to couple NusB to RNAP, as NusB binds to S10 (S. Mason, J. Li, and J.…”

Section: The Role Of Ribosomal Protein S10 In Transcriptionmentioning

confidence: 99%

Assembly of transcription elongation complexes containing the N protein of phage lambda and the Escherichia coli elongation factors NusA, NusB, NusG, and S10.

Mason

Greenblatt²

1991

Genes Dev.

133

126

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The transcription antitermination protein, N, of bacteriophage k; the Escherichia coIi elongation factors NusA, NusB, ribosomal protein $10, and NusG; and a DNA template containing a knut (N-ututilization) site are necessary and sufficient for the highly cooperative formation in vitro of stable transcription complexes containing all five elongation factors. Mutations in the nut site, NusA, or the [bsubunit of RNA polymerase (RNAP) that impair antitermination in vivo also abolish the assembly of a stable complex containing the antitermination factors in vitro. The effects of RNAP mutations on assembly imply that the antitermination factors assemble on the surface of RNAP. We have shown previously that NusA binds directly to transcribing RNAP (K a -10 7 M-l); Ka = association constant and we show here that S10 also binds directly and specifically to RNAP with an apparent K~ of 10 6 M -1. These observations led to a model for the ordered assembly of the N-modified transcription complex.

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Evidence that ribosomal protein S10 itself is a cellular component necessary for transcription antitermination by phage lambda N protein.

Cited by 29 publications

References 34 publications

A New Look at Bacteriophage λ Genetic Networks

A New Look at Bacteriophage λ Genetic Networks

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

Assembly of transcription elongation complexes containing the N protein of phage lambda and the Escherichia coli elongation factors NusA, NusB, NusG, and S10.

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