Escherichia coli K-12 substr. MG1655 Protein: NusB-S10 complex

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].

Reviews: [Weisberg08, Burmann11, Santangelo11]

Citations: [Mogridge98, Das08, Burmann10]

Molecular Weight: 27.0 kD (experimental) [Luttgen02]

Relationship Links: PDB:Structure:3D3B, PDB:Structure:3D3C

Gene-Reaction Schematic

Gene-Reaction Schematic

GO Terms:
Molecular Function:
Inferred from experimentGO:0001068 - transcription regulatory region RNA binding [Luttgen02]
Inferred from experimentGO:0003723 - RNA binding [Luttgen02]

Created 02-Jul-2010 by Keseler I, SRI International

Subunit of NusB-S10 complex: 30S ribosomal subunit protein S10

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]

GO Terms:
Biological Process:
Inferred from experimentGO:0031564 - transcription antitermination [Luo08]
Inferred by computational analysisGO:0006412 - translation [GOA06, GOA01]
Molecular Function:
Inferred from experimentGO:0001072 - transcription antitermination factor activity, RNA binding [Luo08]
Inferred from experimentGO:0005515 - protein binding [Drogemuller15, Burmann12, Burmann10a, Zheng11, Arifuzzaman06, Mason92, Butland05]
Inferred by computational analysisGO:0000049 - tRNA binding [GOA06]
Inferred by computational analysisGO:0003723 - RNA binding [GOA01]
Inferred by computational analysisGO:0003735 - structural constituent of ribosome [GOA01, Gaudet10]
Cellular Component:
Inferred from experimentInferred by computational analysisGO:0005829 - cytosol [DiazMejia09, Ishihama08]
Inferred from experimentGO:0022627 - cytosolic small ribosomal subunit [Hindennach71, Hardy69]
Inferred by computational analysisGO:0005622 - intracellular [GOA01]
Inferred by computational analysisGO:0005840 - ribosome [UniProtGOA11, GOA01]
Inferred by computational analysisGO:0030529 - ribonucleoprotein complex [UniProtGOA11]

MultiFun Terms: cell structureribosomes
information transferprotein relatedribosomal proteins
information transferprotein relatedtranslation
regulationtype of regulationtranscriptional level

Unification Links: DIP:DIP-35797N, EcoliWiki:b3321, Mint:MINT-1283940, ModBase:P0A7R5, PR:PRO_000023863, Pride:P0A7R5, Protein Model Portal:P0A7R5, RefSeq:NP_417780, SMR:P0A7R5, UniProt:P0A7R5

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"

Reviews: [Squires00, Weisberg08, Roberts10]

Citations: [Das08, Sullivan92]

Essentiality data for rpsJ knockouts:

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB LennoxNo 37 Aerobic 7   No [Baba06, Comment 1]

Subunit of NusB-S10 complex: transcription antitermination protein NusB

Synonyms: SsyB, SsaD, NusB

Gene: nusB Accession Numbers: EG10666 (EcoCyc), b0416, ECK0410

Locations: cytosol

Sequence Length: 139 AAs

Molecular Weight: 15.689 kD (from nucleotide sequence)

Molecular Weight: 14 kD (experimental) [Swindle81]

pI: 7.3 [Maekawa85]

GO Terms:
Biological Process:
Inferred by computational analysisGO:0006351 - transcription, DNA-templated [UniProtGOA11]
Inferred by computational analysisGO:0006353 - DNA-templated transcription, termination [UniProtGOA11, GOA06, GOA01]
Inferred by computational analysisGO:0006355 - regulation of transcription, DNA-templated [UniProtGOA11, GOA01]
Molecular Function:
Inferred from experimentGO:0005515 - protein binding [Arifuzzaman06, Butland05, Mason92]
Inferred by computational analysisGO:0003723 - RNA binding [UniProtGOA11, GOA01]
Cellular Component:
Inferred from experimentInferred by computational analysisGO:0005829 - cytosol [DiazMejia09, Ishihama08, LopezCampistrou05]

MultiFun Terms: information transferRNA relatedTranscription related

Unification Links: DIP:DIP-48254N, EcoliWiki:b0416, Mint:MINT-1220894, ModBase:P0A780, PR:PRO_000023446, Pride:P0A780, Protein Model Portal:P0A780, RefSeq:NP_414950, SMR:P0A780, UniProt:P0A780

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].

NusB has also been implicated in termination, including the enhancement of Rho-dependent termination in at least one case [Ward81, Carlomagno01].

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].

NusB is present at about 50-80% of the abundance of the core RNA polymerase [Swindle88]. In the absence of NusB, RNA transcript levels are lower [Miyashita82].

The structural basis of discernment between boxA RNA and its cognate DNA by NusB has been examined [Muhlberger03].

Citations: [Strauch81]

Essentiality data for nusB knockouts:

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB LennoxIndeterminate 37 Aerobic 7   Low [Bubunenko07]
Yes [Baba06, Comment 2]
M9 medium with 1% glycerolYes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucoseYes 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

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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

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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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GOA01: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

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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

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Torres04: Torres M, Balada JM, Zellars M, Squires C, Squires CL (2004). "In vivo effect of NusB and NusG on rRNA transcription antitermination." J Bacteriol 186(5);1304-10. PMID: 14973028

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
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