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PMID:9362499

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Citation

Safo, MK, Yang, WZ, Corselli, L, Cramton, SE, Yuan, HS and Johnson, RC (1997) The transactivation region of the fis protein that controls site-specific DNA inversion contains extended mobile beta-hairpin arms. EMBO J. 16:6860-73

Abstract

The Fis protein regulates site-specific DNA inversion catalyzed by a family of DNA invertases when bound to a cis-acting recombinational enhancer. As is often found for transactivation domains, previous crystal structures have failed to resolve the conformation of the N-terminal inversion activation region within the Fis dimer. A new crystal form of a mutant Fis protein now reveals that the activation region contains two beta-hairpin arms that protrude over 20 A from the protein core. Saturation mutagenesis identified the regulatory and structurally important amino acids. The most critical activating residues are located near the tips of the beta-arms. Disulfide cross-linking between the beta-arms demonstrated that they are highly flexible in solution and that efficient inversion activation can occur when the beta-arms are covalently linked together. The emerging picture for this regulatory motif is that contacts with the recombinase at the tip of the mobile beta-arms activate the DNA invertase in the context of an invertasome complex.

Links

PubMed PMC1170289 Online version:10.1093/emboj/16.22.6860

Keywords

Amino Acid Sequence; Carrier Proteins/chemistry; Carrier Proteins/genetics; Carrier Proteins/metabolism; Computer Simulation; Crystallography, X-Ray; Cysteine/chemistry; DNA Mutational Analysis; DNA Nucleotidyltransferases/metabolism; Factor For Inversion Stimulation Protein; Integration Host Factors; Models, Molecular; Molecular Sequence Data; Oxidation-Reduction; Protein Structure, Secondary; Recombination, Genetic; Trans-Activators/chemistry; Trans-Activators/genetics; Trans-Activators/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ECOLI:FIS

enables

GO:0001046: core promoter sequence-specific DNA binding

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

ECOLI:FIS

involved_in

GO:0006351: transcription, DNA-templated

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:RPOH

enables

GO:0001046: core promoter sequence-specific DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ECOLI:RPOH

involved_in

GO:0006351: transcription, DNA-templated

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:FIS

enables

GO:0001216: DNA-binding transcription activator activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

ECOLI:FIS

involved_in

GO:0001121: bacterial transcription

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:FIS

enables

GO:0044374: sequence-specific DNA binding, bending

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

ECOLI:FIS

involved_in

GO:0032359: provirus excision

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:FIS

enables

GO:0042803: protein homodimerization activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

ECOLI:FIS

part_of

GO:0031421: invertasome

ECO:0000315: mutant phenotype evidence used in manual assertion

C

Seeded From UniProt

complete

ECOLI:FIS

enables

GO:0000985: bacterial-type RNA polymerase core promoter sequence-specific DNA binding

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

ECOLI:FIS

involved_in

GO:0045911: positive regulation of DNA recombination

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:RPOH

part_of

GO:0031421: invertasome

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

ECOLI:RPOH

enables

GO:0009009: site-specific recombinase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ECOLI:RPOH

involved_in

GO:0006310: DNA recombination

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:RPOH

involved_in

GO:0001121: bacterial transcription

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:RPOH

enables

GO:0001216: DNA-binding transcription activator activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ECOLI:RPOH

enables

GO:0000985: bacterial-type RNA polymerase core promoter sequence-specific DNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

Notes

See also

References

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