P0ACJ8 · CRP_ECOLI
- ProteinDNA-binding transcriptional dual regulator CRP
- Genecrp
- StatusUniProtKB reviewed (Swiss-Prot)
- Organism
- Amino acids210 (go to sequence)
- Protein existenceEvidence at protein level
- Annotation score5/5
Function
function
A global transcription regulator, which plays a major role in carbon catabolite repression (CCR) as well as other processes. Binds cyclic AMP (cAMP) which allosterically activates DNA binding (to consensus sequence 5'-AAATGTGATCTAGATCACATTT-3') to directly regulate the transcription of about 300 genes in about 200 operons and indirectly regulate the expression of about half the genome. There are 3 classes of CRP promoters; class I promoters have a single CRP-binding site upstream of the RNA polymerase (RNAP)-binding site, whereas in class II promoters the single CRP- and RNAP-binding site overlap, CRP making multiple contacts with RNAP. Class III promoters require multiple activator molecules, including at least one CRP dimer. CRP can act as an activator, repressor, coactivator or corepressor. Induces a severe bend in DNA (about 87 degrees), bringing upstream promoter elements into contact with RNAP. Acts as a negative regulator of its own synthesis as well as for adenylate cyclase (cyaA), which generates cAMP. High levels of active CRP are detrimental to growth (PubMed:16260780).
In CCR it prevents expression of genes involved in catabolism of nonpreferred carbon sources when glucose is present. CCR involves cAMP, adenylate cyclase (cyaA), CRP and the EIIA-Glc component of the PTS (crr). In the presence of glucose, EIIA-Glc is dephosphorylated, and does not activate adenylate cyclase, leading to reduced cAMP and thus decreased CRP activity. When glucose is absent cAMP binds to CRP, upregulating expression of genes involved in the consumption of non-glucose carbon sources. Also plays a role in many other processes (see PubMed:22573269).
In CCR it prevents expression of genes involved in catabolism of nonpreferred carbon sources when glucose is present. CCR involves cAMP, adenylate cyclase (cyaA), CRP and the EIIA-Glc component of the PTS (crr). In the presence of glucose, EIIA-Glc is dephosphorylated, and does not activate adenylate cyclase, leading to reduced cAMP and thus decreased CRP activity. When glucose is absent cAMP binds to CRP, upregulating expression of genes involved in the consumption of non-glucose carbon sources. Also plays a role in many other processes (see PubMed:22573269).
Among the targets of CRP is the spf gene which encodes the dual-function sRNA Spot 42; spf expression is repressed by CRP (PubMed:2454912).
Spot 42 base pairs with and represses expression from a number of mRNAs encoding proteins involved in the uptake and catabolism of nonpreferred carbon sources. CRP and Spot 42 coregulate at least 14 operons involved in cellular metabolism, forming a coherent feedforward loop (PubMed:21292161).
Interaction with SpfP, encoded within the Spot 42 RNA, reinforces this feedforward loop (PubMed:35239441).
Spot 42 base pairs with and represses expression from a number of mRNAs encoding proteins involved in the uptake and catabolism of nonpreferred carbon sources. CRP and Spot 42 coregulate at least 14 operons involved in cellular metabolism, forming a coherent feedforward loop (PubMed:21292161).
Interaction with SpfP, encoded within the Spot 42 RNA, reinforces this feedforward loop (PubMed:35239441).
Miscellaneous
Binds 2 cAMP; cAMP 1 is in the anti conformation, while cAMP 2 is in the syn conformation.
Activity regulation
In the apo-form the DNA-binding helices form a rigid body in which their DNA recognitions helices are buried. cAMP binding causes a coil-to-helix transition, stabilizing the active DNA binding conformation by reorienting and elongating these helices, which precludes a return to the inactive state (PubMed:19359484, PubMed:19805344).
Interaction with SpfP blocks activation of some, but not all, CRP-dependent genes (PubMed:35239441).
Interaction with SpfP blocks activation of some, but not all, CRP-dependent genes (PubMed:35239441).
Features
Showing features for binding site, site, dna binding.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Binding site | 57-63 | 3',5'-cyclic AMP 1 (UniProtKB | ChEBI) | ||||
Sequence: GKEMILS | ||||||
Binding site | 72-74 | 3',5'-cyclic AMP 1 (UniProtKB | ChEBI) | ||||
Sequence: GEL | ||||||
Binding site | 83-84 | 3',5'-cyclic AMP 1 (UniProtKB | ChEBI) | ||||
Sequence: RS | ||||||
Site | 97 | Activating region 2 (AR2); probably contacts the N-terminus of RpoA | ||||
Sequence: E | ||||||
Site | 102 | Activating region 2 (AR2); probably contacts the N-terminus of RpoA | ||||
Sequence: K | ||||||
Binding site | 128-129 | 3',5'-cyclic AMP 1 (UniProtKB | ChEBI) | ||||
Sequence: TS | ||||||
Binding site | 136-137 | 3',5'-cyclic AMP 2 (UniProtKB | ChEBI) | ||||
Sequence: AF | ||||||
Binding site | 171-181 | 3',5'-cyclic AMP 2 (UniProtKB | ChEBI) | ||||
Sequence: QEIGQIVGCSR | ||||||
DNA binding | 180-186 | H-T-H motif | ||||
Sequence: SRETVGR |
GO annotations
Aspect | Term | |
---|---|---|
Cellular Component | cytosol | |
Cellular Component | protein-DNA complex | |
Molecular Function | cAMP binding | |
Molecular Function | DNA binding, bending | |
Molecular Function | DNA-binding transcription factor activity | |
Molecular Function | identical protein binding | |
Molecular Function | minor groove of adenine-thymine-rich DNA binding | |
Molecular Function | sequence-specific DNA binding | |
Biological Process | carbon catabolite repression of transcription | |
Biological Process | DNA-templated transcription | |
Biological Process | negative regulation of DNA-templated transcription | |
Biological Process | positive regulation of DNA-templated transcription |
Keywords
- Molecular function
- Biological process
- Ligand
Enzyme and pathway databases
Names & Taxonomy
Protein names
- Recommended nameDNA-binding transcriptional dual regulator CRP
- Alternative names
Gene names
Organism names
- Organism
- Strains
- Taxonomic lineageBacteria > Pseudomonadota > Gammaproteobacteria > Enterobacterales > Enterobacteriaceae > Escherichia
Accessions
- Primary accessionP0ACJ8
- Secondary accessions
Proteomes
Subcellular Location
UniProt Annotation
GO Annotation
Phenotypes & Variants
Disruption phenotype
Not essential (on rich medium), greatly increased levels of cAMP. Eliminates the NaCl sensitivity of an rnlA deletion mutant.
Features
Showing features for mutagenesis.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Mutagenesis | 20 | Decreased transcription activation at class II promoters, decreased interaction with RNAP, binds DNA. | ||||
Sequence: H → A, L, or Y | ||||||
Mutagenesis | 22 | Decreased transcription activation at class II promoters, decreased interaction with RNAP, binds DNA. | ||||
Sequence: H → A or L | ||||||
Mutagenesis | 51 | Prevents SpfP from blocking galactose operon induction by CRP. | ||||
Sequence: L → M | ||||||
Mutagenesis | 53 | Increased activation at class II promoters, increased interaction with RNAP. | ||||
Sequence: K → N | ||||||
Mutagenesis | 54-56 | 80% reduction in activation of class II promoters; 95% loss when associated with A-59. | ||||
Sequence: DEE → AAA | ||||||
Mutagenesis | 59 | 45% reduction in activation of class II promoters; 95% loss when associated with AAA-54-56. | ||||
Sequence: E → A | ||||||
Mutagenesis | 59 | Reduction in activation of class II promoters. | ||||
Sequence: E → G or K | ||||||
Mutagenesis | 62 | Prevents SpfP from blocking galactose operon induction by CRP. | ||||
Sequence: L → F | ||||||
Mutagenesis | 63 | Enhanced cAMP-binding, enhanced transcription. | ||||
Sequence: S → A | ||||||
Mutagenesis | 83 | Loss of cAMP-binding. | ||||
Sequence: R → L | ||||||
Mutagenesis | 84 | No modification of cAMP-binding. | ||||
Sequence: S → A or K | ||||||
Mutagenesis | 84 | Prevents SpfP from blocking galactose operon induction by CRP. | ||||
Sequence: S → R | ||||||
Mutagenesis | 91 | Growth defect on glucose and galactose. | ||||
Sequence: T → S | ||||||
Mutagenesis | 97 | Increased transcription activation at class II promoters, binds DNA. | ||||
Sequence: E → A | ||||||
Mutagenesis | 102 | Disrupts AR2. No activation of class II promoters, decreased interaction with RNAP, binds DNA. | ||||
Sequence: K → E | ||||||
Mutagenesis | 128 | No modification of cAMP-binding. | ||||
Sequence: T → A | ||||||
Mutagenesis | 128-129 | Constitutively active at class I and II promoters in the absence of cAMP, binds DNA almost as well in the absence as in the presence of cAMP. Binds cAMP normally. | ||||
Sequence: TS → LI | ||||||
Mutagenesis | 129 | Reduced DNA-binding; no modification of cAMP-binding. | ||||
Sequence: S → A | ||||||
Mutagenesis | 139 | Some stabilization of an inactive (apo-) form. Decreased affinity for DNA, normal subunit association. | ||||
Sequence: D → L | ||||||
Mutagenesis | 157 | Decreased transcription activation (6-29%), binds DNA. | ||||
Sequence: A → D or P | ||||||
Mutagenesis | 159 | Decreased transcription activation (15-87%) at class I and II promoters, binds DNA. | ||||
Sequence: T → A, I, N, S, or V | ||||||
Mutagenesis | 160 | Disrupts AR1. Decreased transcription activation (3-45%) at class I and II promoters, binds DNA. | ||||
Sequence: H → A, K, L, N, P, Q, R, or Y | ||||||
Mutagenesis | 163 | Decreased transcription activation (2-62%) at class I and II promoters, binds DNA. | ||||
Sequence: G → A, C, D, R, S, or V | ||||||
Mutagenesis | 180 | Prevents SpfP from blocking galactose operon induction by CRP. | ||||
Sequence: S → Y | ||||||
Mutagenesis | 181 | Suppresses DNA-binding. | ||||
Sequence: R → K | ||||||
Mutagenesis | 181 | Suppresses DNA-binding. | ||||
Sequence: R → L | ||||||
Mutagenesis | 186 | No modification of DNA-binding. | ||||
Sequence: R → K | ||||||
Mutagenesis | 186 | Marginally reduced DNA-binding. | ||||
Sequence: R → L |
Chemistry
PTM/Processing
Features
Showing features for initiator methionine, chain, modified residue.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Initiator methionine | 1 | Removed | ||||
Sequence: M | ||||||
Chain | PRO_0000100144 | 2-210 | DNA-binding transcriptional dual regulator CRP | |||
Sequence: VLGKPQTDPTLEWFLSHCHIHKYPSKSTLIHQGEKAETLYYIVKGSVAVLIKDEEGKEMILSYLNQGDFIGELGLFEEGQERSAWVRAKTACEVAEISYKKFRQLIQVNPDILMRLSAQMARRLQVTSEKVGNLAFLDVTGRIAQTLLNLAKQPDAMTHPDGMQIKITRQEIGQIVGCSRETVGRILKMLEDQNLISAHGKTIVVYGTR | ||||||
Modified residue | 101 | N6-acetyllysine | ||||
Sequence: K |
Keywords
- PTM
Proteomic databases
PTM databases
Expression
Interaction
Subunit
Homodimer, which upon binding cAMP is able to bind DNA. AR1 of the upstream subunit binds to the C-terminus of RNAP subunit RpoA, AR2 of the downstream subunit binds to the N-terminus of RpoA while AR3 binds to sigma-70 (rpoD) (PubMed:11124031, PubMed:12202833, PubMed:1646077, PubMed:1653449, PubMed:19359484, PubMed:19805344, PubMed:19903881, PubMed:2828639, PubMed:2839152, PubMed:6286624, PubMed:8757802, PubMed:8978616, PubMed:9096308, Ref.44). Interacts with SpfP (PubMed:35239441).
Binary interactions
Type | Entry 1 | Entry 2 | Number of experiments | Intact | |
---|---|---|---|---|---|
BINARY | P0ACJ8 | crp P0ACJ8 | 5 | EBI-547513, EBI-547513 | |
BINARY | P0ACJ8 | nusG P0AFG0 | 2 | EBI-547513, EBI-369628 | |
BINARY | P0ACJ8 | priC P23862 | 2 | EBI-547513, EBI-1117383 | |
BINARY | P0ACJ8 | rplL P0A7K2 | 2 | EBI-547513, EBI-543702 | |
BINARY | P0ACJ8 | yacL P0A8E5 | 4 | EBI-547513, EBI-554965 |
Protein-protein interaction databases
Structure
Family & Domains
Features
Showing features for region, domain.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Region | 20-22 | Activating region 2 (AR2); probably contacts the N-terminus of RpoA | ||||
Sequence: HIH | ||||||
Region | 53-59 | Activating region 3 (AR3); probably contacts sigma-70 (RpoD) | ||||
Sequence: KDEEGKE | ||||||
Domain | 138-210 | HTH crp-type | ||||
Sequence: LDVTGRIAQTLLNLAKQPDAMTHPDGMQIKITRQEIGQIVGCSRETVGRILKMLEDQNLISAHGKTIVVYGTR | ||||||
Region | 154-163 | Activating region 1 (AR1); probably contacts the C-terminus of RpoA | ||||
Sequence: QPDAMTHPDG |
Domain
The N-terminal domain binds cAMP and is responsible for homodimerization, while the C-terminal domain binds DNA when cAMP is bound.
Phylogenomic databases
Family and domain databases
Sequence
- Sequence statusComplete
- Length210
- Mass (Da)23,640
- Last updated1986-07-21 v1
- ChecksumDCBC24FA46C61B3D
Features
Showing features for sequence conflict.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Sequence conflict | 29 | in Ref. 4; BAE77933 | ||||
Sequence: T → K |
Keywords
- Technical term
Sequence databases
Nucleotide Sequence | Protein Sequence | Molecule Type | Status | |
---|---|---|---|---|
J01598 EMBL· GenBank· DDBJ | AAA23601.1 EMBL· GenBank· DDBJ | Genomic DNA | ||
U18997 EMBL· GenBank· DDBJ | AAA58154.1 EMBL· GenBank· DDBJ | Genomic DNA | ||
U00096 EMBL· GenBank· DDBJ | AAC76382.1 EMBL· GenBank· DDBJ | Genomic DNA | ||
AP009048 EMBL· GenBank· DDBJ | BAE77933.1 EMBL· GenBank· DDBJ | Genomic DNA |