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 [Burmann10]. 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 [Das84, 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]|
Altieri00: Altieri AS, Mazzulla MJ, Horita DA, Coats RH, Wingfield PT, Das A, Court DL, Byrd RA (2000). "The structure of the transcriptional antiterminator NusB from Escherichia coli." Nat Struct Biol 7(6);470-4. PMID: 10881193
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
Burmann10a: 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
Das84: Das A, Wolska K (1984). "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 38(1);165-73. PMID: 6088061
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
Friedman81: Friedman DI, Schauer AT, Baumann MR, Baron LS, Adhya SL (1981). "Evidence that ribosomal protein S10 participates in control of transcription termination." Proc Natl Acad Sci U S A 78(2);1115-8. PMID: 6453343
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
Luo08: Luo X, Hsiao HH, Bubunenko M, Weber G, Court DL, Gottesman ME, Urlaub H, Wahl MC (2008). "Structural and functional analysis of the E. coli NusB-S10 transcription antitermination complex." Mol Cell 32(6);791-802. PMID: 19111659
Luttgen02: Luttgen H, Robelek R, Muhlberger R, Diercks T, Schuster SC, Kohler P, Kessler H, Bacher A, Richter G (2002). "Transcriptional regulation by antitermination. Interaction of RNA with NusB protein and NusB/NusE protein complex of Escherichia coli." J Mol Biol 316(4);875-85. PMID: 11884128
Mason91: Mason SW, Greenblatt J (1991). "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 5(8);1504-12. PMID: 1831176
Mason92: Mason SW, Li J, Greenblatt J (1992). "Direct interaction between two Escherichia coli transcription antitermination factors, NusB and ribosomal protein S10." J Mol Biol 223(1);55-66. PMID: 1731086
Miyashita82: Miyashita T, Kano Y, Kuroki K, Ishii S, Imamoto F (1982). "In vivo evidence for nusA and nusB gene function in general transcription of the Escherichia coli genome." Biken J 25(3);121-30. PMID: 6187334
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
Muhlberger03: Muhlberger R, Robelek R, Eisenreich W, Ettenhuber C, Sinner EK, Kessler H, Bacher A, Richter G (2003). "RNA DNA discrimination by the antitermination protein NusB." J Mol Biol 327(5);973-83. PMID: 12662923
Powers88: Powers T, Stern S, Changchien LM, Noller HF (1988). "Probing the assembly of the 3' major domain of 16 S rRNA. Interactions involving ribosomal proteins S2, S3, S10, S13 and S14." J Mol Biol 201(4);697-716. PMID: 2459390
Sharrock85: Sharrock RA, Gourse RL, Nomura M (1985). "Defective antitermination of rRNA transcription and derepression of rRNA and tRNA synthesis in the nusB5 mutant of Escherichia coli." Proc Natl Acad Sci U S A 82(16);5275-9. PMID: 3161080
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
Zheng11: Zheng C, Yang L, Hoopmann MR, Eng JK, Tang X, Weisbrod CR, Bruce JE (2011). "Cross-linking measurements of in vivo protein complex topologies." Mol Cell Proteomics 10(10);M110.006841. PMID: 21697552
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