Crp of Streptomyces coelicolor is the third transcription factor of the large CRP-FNR superfamily able to bind cAMP.
The chromosomal inactivation of the unique transcription factor of Streptomyces coelicolor that displays a cyclic-nucleotide-binding domain, Crp(Sco), led to a germination-defective phenotype similar to the mutant of the adenylate cyclase gene (cya) unable to produce cAMP. By means of cAMP affinity chromatography we demonstrate the specific cAMP-binding ability of Crp(Sco), which definitely demonstrate that a Cya/cAMP/Crp system is used to trigger germination in S. coelicolor. However, electromobility shift assays with the purified Crp(Sco)-cAMP complex and the CRP-like cis-acting element of its own promoter failed. Moreover, we were unable to complement an Escherichia coli crp mutant in trans with Crp(Sco). The fact that Vfr from Pseudomonas aeruginosa and GlxR from Corynebacterium glutamicum could complement such an E. coli mutant suggests that the way Crp(Sco) interacts with DNA should mechanistically differ from its most similar members. This hypothesis was further supported by homology modelling of Crp(Sco) that confirmed an unusual organisation of the DNA-binding domain compared to the situation observed in Crp(Eco).
Derouaux
Adeline
A
Centre d'Ingénierie des Protéines, Université de Liège, Institut de Chimie B6a, B-4000 Liège, Belgium.
Dehareng
Dominique
D
Lecocq
Elke
E
Halici
Serkan
S
Nothaft
Harald
H
Giannotta
Fabrizio
F
Moutzourelis
Georgios
G
Dusart
Jean
J
Devreese
Bart
B
Titgemeyer
Fritz
F
Van Beeumen
Jozef
J
Rigali
Sébastien
S
D000595:Amino Acid Sequence; D001665:Binding Sites; D003198:Computer Simulation; D000242:Cyclic AMP / Q000737:chemistry* / Q000378:metabolism*; D002373:Cyclic AMP Receptor Protein / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000145:classification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D048372:Streptomyces coelicolor / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism
D002373:Cyclic AMP Receptor Protein; D011994:Recombinant Proteins; D014157:Transcription Factors; D000242:Cyclic AMP