Synonyms: NusB-NusE complex
Subunit composition of
NusB-S10 complex = [RpsJ][NusB]
30S ribosomal subunit protein S10 = RpsJ (extended summary available)
transcription antitermination protein NusB = NusB (extended summary available)
The NusB-S10(NusE) complex is involved in modulating transcription. It was first discovered for its role in antitermination of phage λ early transcription [Friedman81] and was later shown to be required for antitermination in rRNA operons and, surprisingly, transcription termination mediated by the Nun protein of phage HK022.
NusB and S10 form a heterodimer [Mason92] that specifically binds transcripts containing the BoxA sequence [Nodwell93, Luttgen02]. Detailed analysis of the assembly pathway for the BoxA-containing core antitermination complex has been performed [Greive05]. S10 was shown to bind RNA non-specifically and increases the affinity of NusB for boxA RNA [Greive05].
An S10 mutant that lacks the extended ribosome binding loop (aa 46-67, S10Δloop) retains its regulatory function in transcription termination, but can not perform the essential function of S10, i.e. likely its role in the ribosome/translation [Luo08]. A crystal structure of the NusB-S10Δloop complex has been solved [Luo08]. UV-induced protein-RNA crosslinking of NusB and S10 shows a continuous BoxA binding surface involving both proteins and suggests that NusB stabilizes a BoxA-binding conformation of S10 [Luo08]. Overexpression of S10 in a strain lacking NusB rescues transcription antitermination, indicating that S10 is the critical component of the NusB-S10 complex, while NusB appears to recruit S10 to the BoxA binding site [Luo08].
Molecular Weight: 27.0 kD (experimental) [Luttgen02]
Synonyms: NusE, RpsJ
|Gene:||rpsJ||Accession Numbers: EG10909 (EcoCyc), b3321, ECK3308|
Locations: cytosol, ribosome
Sequence Length: 103 AAs
Molecular Weight: 11.736 kD (from nucleotide sequence)
Molecular Weight: 12.0 kD (experimental) [Mason92]
|MultiFun Terms:||cell structure → ribosomes|
|information transfer → protein related → ribosomal proteins|
|information transfer → protein related → translation|
|regulation → type of regulation → transcriptional level|
Relationship Links: InterPro:IN-FAMILY:IPR001848, InterPro:IN-FAMILY:IPR018268, InterPro:IN-FAMILY:IPR027486, Panther:IN-FAMILY:PTHR11700, PDB:Structure:1M5G, PDB:Structure:2KVQ, PDB:Structure:2YKR, PDB:Structure:3D3B, PDB:Structure:3D3C, PDB:Structure:3IMQ, PDB:Structure:3J9Y, PDB:Structure:3W1Y, PDB:Structure:4A2I, PDB:Structure:4ADV, PDB:Structure:4U1U, PDB:Structure:4U1V, PDB:Structure:4U20, PDB:Structure:4U24, PDB:Structure:4U25, PDB:Structure:4U26, PDB:Structure:4U27, PDB:Structure:4V6K, PDB:Structure:4V6L, PDB:Structure:4V6M, PDB:Structure:4V6N, PDB:Structure:4V6O, PDB:Structure:4V6P, PDB:Structure:4V6Q, PDB:Structure:4V6R, PDB:Structure:4V6S, PDB:Structure:4V6T, PDB:Structure:4V6V, PDB:Structure:4V6Y, PDB:Structure:4V6Z, PDB:Structure:4V9C, PDB:Structure:4V9D, PDB:Structure:4V9O, PDB:Structure:4V9P, PDB:Structure:4V47, PDB:Structure:4V48, PDB:Structure:4V4H, PDB:Structure:4V4Q, PDB:Structure:4V4V, PDB:Structure:4V4W, PDB:Structure:4V50, PDB:Structure:4V52, PDB:Structure:4V53, PDB:Structure:4V54, PDB:Structure:4V55, PDB:Structure:4V56, PDB:Structure:4V57, PDB:Structure:4V5B, PDB:Structure:4V5H, PDB:Structure:4V5Y, PDB:Structure:4V64, PDB:Structure:4V65, PDB:Structure:4V66, PDB:Structure:4V69, PDB:Structure:4V6C, PDB:Structure:4V6D, PDB:Structure:4V6E, PDB:Structure:4V70, PDB:Structure:4V71, PDB:Structure:4V72, PDB:Structure:4V73, PDB:Structure:4V74, PDB:Structure:4V75, PDB:Structure:4V76, PDB:Structure:4V77, PDB:Structure:4V78, PDB:Structure:4V79, PDB:Structure:4V7A, PDB:Structure:4V7B, PDB:Structure:4V7C, PDB:Structure:4V7D, PDB:Structure:4V7I, PDB:Structure:4V7S, PDB:Structure:4V7T, PDB:Structure:4V7U, PDB:Structure:4V7V, PDB:Structure:4V85, PDB:Structure:4V89, PDB:Structure:4WF1, PDB:Structure:4WWW, PDB:Structure:4YBB, PDB:Structure:5AFI, Pfam:IN-FAMILY:PF00338, Prints:IN-FAMILY:PR00971, Prosite:IN-FAMILY:PS00361
The S10 protein (NusE) is a component of the 30S subunit of the ribosome. From within its location in the ribosome, S10 plays a role in linking transcription and translation; in a separate complex with NusB, it plays a role in regulating transcription antitermination.
S10 can be crosslinked to tRNA in the ribosomal P site [Riehl82] and may contact 16S rRNA in two separate domains [Powers88]. Mutations in S10 confer resistance to the antibiotic tigecycline [Beabout15].
As part of the ribosome, S10 also interacts with several non-ribosomal proteins. The NusG protein interacts with S10 and may thereby physically link transcription and translation [Burmann10a]. Interaction of the NTD of RfaH with its ops DNA target releases its CTD, which then switches to a conformation that can interact with S10 and activate translation [Burmann12].
Outside of the ribosome, S10 is involved in the regulation of transcription termination [Das84a, Warren84, Das85]. S10 forms a heterodimer with NusB [Mason92] which can bind to the boxA sequence of rrn operons [Nodwell93, Luttgen02]. Detailed analysis of the assembly pathway for the boxA-containing core antitermination complex has been performed. S10 was shown to bind RNA non-specifically and increases the affinity of NusB for boxA RNA [Greive05]. Because overexpression of S10 in a strain lacking NusB rescues transcription antitermination, S10 is the critical component of the NusB-S10 complex, while NusB appears to serve as a loading factor [Luo08]. An S10 mutant that lacks the extended ribosome binding loop (aa 46-67, S10Δloop) retains its regulatory function in transcription termination, but can not perform the essential function of S10, i.e. likely its role in the ribosome/translation. The interactions of S10 with the ribosome or with NusB are mutually exclusive [Luo08]. Within the NusB-S10 complex, S10 can interact with RpoB, the β subunit of RNA polymerase [Mason91, Drogemuller15].
S10's extraribosomal function was first discovered in the context of bacteriophage λ biology. The nusE71 mutation in the rpsJ gene was shown to affect regulation of transcription termination by the bacteriophage λ antiterminator N [Friedman81]. The nusE71 allele is a point mutation, changing a single amino acid, Ala86, to Asp. It is the only rpsJ allele known to have a Nus- phenotype [Court95]. The nusE71 mutation may be specific for λ antitermination; it appears to have no effect on boxA-mediated increase in the rate of transcription of rrn operons [Zellars99].
NusE: "N utilization substance E"
|Growth Medium||Growth?||T (°C)||O2||pH||Osm/L||Growth Observations|
|LB Lennox||No||37||Aerobic||7||No [Baba06, Comment 1]|
Synonyms: SsyB, SsaD, NusB
|Gene:||nusB||Accession Numbers: EG10666 (EcoCyc), b0416, ECK0410|
Sequence Length: 139 AAs
Molecular Weight: 15.689 kD (from nucleotide sequence)
Molecular Weight: 14 kD (experimental) [Swindle81]
pI: 7.3 [Maekawa85]
|MultiFun Terms:||information transfer → RNA related → Transcription related|
Relationship Links: InterPro:IN-FAMILY:IPR006027, InterPro:IN-FAMILY:IPR011605, Panther:IN-FAMILY:PTHR11078, PDB:Structure:1EY1, PDB:Structure:3D3B, PDB:Structure:3D3C, PDB:Structure:3IMQ, Pfam:IN-FAMILY:PF01029
Transcription antitermination protein NusB is involved in antitermination in the transcription of a number of genes. In particular, NusB is required for proper transcription of the ribosomal RNA (rRNA) genes.
NusB is required for antitermination in lambda phage, as reviewed in [Das92].
NusB is involved in antitermination in the cell [Kuroki82]. NusB is strictly required for antitermination and transcription of rRNA genes [Sharrock85, Zellars99, Torres04, Quan05]. This antitermination role requires weak initial binding of a boxA sequence on the nascent RNA by NusB, followed by stabilizing binding by 30S ribosomal subunit protein S10 [Greive05, Luttgen02, Nodwell93, Mason92].
A number of structural analyses have been carried out on NusB. It is active as a monomer [Swindle88]. NMR studies of NusB indicate that it is entirely alpha-helical, with six or seven helices [Berglechner97, Altieri97, Altieri00].
The structural basis of discernment between boxA RNA and its cognate DNA by NusB has been examined [Muhlberger03].
|Growth Medium||Growth?||T (°C)||O2||pH||Osm/L||Growth Observations|
Yes [Baba06, Comment 2]
|M9 medium with 1% glycerol||Yes||37||Aerobic||7.2||0.35||Yes [Joyce06, Comment 3]|
|MOPS medium with 0.4% glucose||Yes||37||Aerobic||7.2||0.22||Yes [Baba06, Comment 2]|
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Altieri97: Altieri AS, Mazzulla MJ, Zhou H, Costantino N, Court DL, Byrd RA (1997). "Sequential assignments and secondary structure of the RNA-binding transcriptional regulator NusB." FEBS Lett 415(2);221-6. PMID: 9351000
Arifuzzaman06: Arifuzzaman M, Maeda M, Itoh A, Nishikata K, Takita C, Saito R, Ara T, Nakahigashi K, Huang HC, Hirai A, Tsuzuki K, Nakamura S, Altaf-Ul-Amin M, Oshima T, Baba T, Yamamoto N, Kawamura T, Ioka-Nakamichi T, Kitagawa M, Tomita M, Kanaya S, Wada C, Mori H (2006). "Large-scale identification of protein-protein interaction of Escherichia coli K-12." Genome Res 16(5);686-91. PMID: 16606699
Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554
Beabout15: Beabout K, Hammerstrom TG, Perez AM, Magalhaes BF, Prater AG, Clements TP, Arias CA, Saxer G, Shamoo Y (2015). "The ribosomal S10 protein is a general target for decreased tigecycline susceptibility." Antimicrob Agents Chemother. PMID: 26124155
Berglechner97: Berglechner F, Richter G, Fischer M, Bacher A, Gschwind RM, Huenges M, Gemmecker G, Kessler H (1997). "Studies on the NusB protein of Escherichia coli--expression and determination of secondary-structure elements by multinuclear NMR spectroscopy." Eur J Biochem 248(2);338-46. PMID: 9346286
Bubunenko07: Bubunenko M, Baker T, Court DL (2007). "Essentiality of ribosomal and transcription antitermination proteins analyzed by systematic gene replacement in Escherichia coli." J Bacteriol 189(7);2844-53. PMID: 17277072
Burmann10: Burmann BM, Luo X, Rosch P, Wahl MC, Gottesman ME (2010). "Fine tuning of the E. coli NusB:NusE complex affinity to BoxA RNA is required for processive antitermination." Nucleic Acids Res 38(1);314-26. PMID: 19854945
Burmann11: Burmann BM, Rosch P (2011). "The role of E. coli Nus-factors in transcription regulation and transcription:translation coupling: From structure to mechanism." Transcription 2(3);130-134. PMID: 21922055
Burmann12: Burmann BM, Knauer SH, Sevostyanova A, Schweimer K, Mooney RA, Landick R, Artsimovitch I, Rosch P (2012). "An α helix to β barrel domain switch transforms the transcription factor RfaH into a translation factor." Cell 150(2);291-303. PMID: 22817892
Butland05: Butland G, Peregrin-Alvarez JM, Li J, Yang W, Yang X, Canadien V, Starostine A, Richards D, Beattie B, Krogan N, Davey M, Parkinson J, Greenblatt J, Emili A (2005). "Interaction network containing conserved and essential protein complexes in Escherichia coli." Nature 433(7025);531-7. PMID: 15690043
Court95: Court DL, Patterson TA, Baker T, Costantino N, Mao X, Friedman DI (1995). "Structural and functional analyses of the transcription-translation proteins NusB and NusE." J Bacteriol 177(9);2589-91. PMID: 7730297
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Das85: Das A, Ghosh B, Barik S, Wolska K (1985). "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 82(12);4070-4. PMID: 2987961
Das92: Das A (1992). "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 174(21);6711-6. PMID: 1400223
DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114
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Greive05: Greive SJ, Lins AF, von Hippel PH (2005). "Assembly of an RNA-protein complex. Binding of NusB and NusE (S10) proteins to boxA RNA nucleates the formation of the antitermination complex involved in controlling rRNA transcription in Escherichia coli." J Biol Chem 280(43);36397-408. PMID: 16109710
Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394
Kuroki82: Kuroki K, Ishii S, Kano Y, Miyashita T, Nishi K, Imamoto F (1982). "Involvement of the nusB gene products in transcription of Escherichia coli tryptophan operon in vitro." Mol Gen Genet 185(2);369-71. PMID: 7045592
LopezCampistrou05: Lopez-Campistrous A, Semchuk P, Burke L, Palmer-Stone T, Brokx SJ, Broderick G, Bottorff D, Bolch S, Weiner JH, Ellison MJ (2005). "Localization, annotation, and comparison of the Escherichia coli K-12 proteome under two states of growth." Mol Cell Proteomics 4(8);1205-9. PMID: 15911532
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Mogridge98: Mogridge J, Mah TF, Greenblatt J (1998). "Involvement of boxA nucleotides in the formation of a stable ribonucleoprotein complex containing the bacteriophage lambda N protein." J Biol Chem 273(7);4143-8. PMID: 9461609
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Warren84: Warren F, Das A (1984). "Formation of termination-resistant transcription complex at phage lambda nut locus: effects of altered translation and a ribosomal mutation." Proc Natl Acad Sci U S A 81(12);3612-6. PMID: 6233610
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