Role of the Cro repressor carboxy terminal domain and flexible dimer linkage in operator and nonspecific DNA binding
| Autor: | Laurent Bracco, Marvin H. Caruthers, Scott J. Eisenbeis, Richard B. Gayle, Graham Beaton, Adrian J. Hubbard |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Mutation
Operator Regions Genetic Operator (biology) Base Sequence biology Stereochemistry Molecular Sequence Data Mutant Repressor DNA DNA-binding domain Lambda phage medicine.disease_cause biology.organism_classification Biochemistry Molecular biology Repressor Proteins chemistry.chemical_compound chemistry medicine Amino Acid Sequence Cloning Molecular Site-directed mutagenesis |
| Zdroj: | Biochemistry. 29:9241-9249 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi00491a019 |
| Popis: | A series of mutations comprising single and multiple substitutions, deletions, and extensions within the carboxy-terminal domain of the bacteriophage lambda Cro repressor have been constructed. These mutations generally affect the affinity of repressor for specific and nonspecific DNA. Additionally, substitution of the carboxy-terminal alanine with several amino acids capable of hydrogen-bonding interactions leads to improved specific binding affinities. A mutation is also described whereby cysteine links the two Cro monomers by a disulfide bond. As a consequence, a significant improvement in nonspecific binding and a concomitant reduction in specific binding are observed with this mutant. These results provide evidence that the carboxy terminus of Cro repressor is an important DNA binding domain and that a flexible connection between the two repressor monomers is a critical factor in modulating the affinity of wild-type repressor for DNA. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|