Amino acid substitution in the C-terminal arm domain of HU-2 results in an enhanced affinity for DNA
| ISSN: | 0378-1119 |
|---|---|
| Přístupová URL adresa: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a4faa5ab276d3a11db2e504acd1d6865 https://pubmed.ncbi.nlm.nih.gov/1427084 |
| Rights: | CLOSED |
| Přírůstkové číslo: | edsair.doi.dedup.....a4faa5ab276d3a11db2e504acd1d6865 |
| Autor: | Goshima Naoki, Kano Yasunobu, Tanaka Hiromitsu, Tanaka Hideyuki, Kohno Kyoko, Yasuzawa Kayoko, Fumio Imamoto |
| Rok vydání: | 1992 |
| Předmět: |
DNA
Bacterial HMG-box Mutant Blotting Western Molecular Sequence Data Biology Frameshift mutation law.invention Bacteriophage mu chemistry.chemical_compound Bacterial Proteins Mutant protein law Genetics Escherichia coli Amino Acid Sequence Amino Acids Site-directed mutagenesis Peptide sequence Base Sequence General Medicine Molecular biology DNA-Binding Proteins chemistry Biochemistry Recombinant DNA Mutagenesis Site-Directed Electrophoresis Polyacrylamide Gel DNA Plasmids Protein Binding |
| Zdroj: | Gene. 121(1) |
| ISSN: | 0378-1119 |
| Popis: | Three mutants of the Escherichia coli hupA gene, encoding the HU-2 protein, were constructed by synthetic oligodeoxyribonucleotide-directed, site-specific mutagenesis on M13mp18 vectors. The resulting HupAN10, HupAN11 and HupAN12 proteins contained Thr59-->Lys, Gln64-->Lys and Asn53-->Arg substitutions, respectively. These amino acid (aa) changes increased the positive charge of the N-terminal half of the two-strand, antiparallel beta-ribbon of the arm structure, which is believed to be a domain for DNA binding. The three mutant proteins bound to DNA more tightly than wild-type HU-2, and their affinities for DNA increased in the order of HupAN10, HupAN11, HupAN12. The mutant proteins showed a slightly increased HU activity for supporting Mu phage development. A mutant HU-2 protein with increased basicity, but with an altered aa sequence in the arm region due to a frameshift mutation, was also constructed. This mutant protein showed a reduced affinity to DNA and was unable to support Mu growth, suggesting that a unique aa sequence of the arm domain, rather than mere basicity of this domain, is required for efficient binding to DNA. |
| Databáze: | OpenAIRE |
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