Hypomorphic mutations in human DNA ligase IV lead to compromised DNA binding efficiency, hydrophobicity and thermal stability
| Autor: | Ramanathan Natesh, Sathees C. Raghavan, Eswar Reddy Maddi |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
chemistry.chemical_classification
DNA ligase Circular dichroism DNA Ligases Chemistry LIG4 syndrome Mutant Wild type Bioengineering DNA medicine.disease Biochemistry DNA Ligase ATP chemistry.chemical_compound Mutation medicine Biophysics Humans Electrophoretic mobility shift assay Protein folding Hydrophobic and Hydrophilic Interactions Molecular Biology Biotechnology |
| Zdroj: | Protein Engineering, Design and Selection. 34 |
| ISSN: | 1741-0134 1741-0126 |
| DOI: | 10.1093/protein/gzab001 |
| Popis: | Studies have shown that Lig4 syndrome mutations in DNA ligase IV (LigIV) are compromised in its function with residual level of double strand break ligation activity in vivo. It was speculated that Lig4 syndrome mutations adversely affect protein folding and stability. Though there are crystal structures of LigIV, there are no reports of crystal structures of Lig4 syndrome mutants and their biophysical characterization to date. Here, we have examined the conformational states, thermal stability, hydrophobicity and DNA binding efficiency of human DNA LigIV wild type and its hypomorphic mutants by far-UV circular dichroism, tyrosine and tryptophan fluorescence, and 1-anilino-8-naphthalene-sulfonate binding, dynamic light scattering, size exclusion chromatography, multi-angle light scattering and electrophoretic mobility shift assay. We show here that LigIV hypomorphic mutants have reduced DNA-binding efficiency, a shift in secondary structure content from the helical to random coil, marginal reduction in their thermal stability and increased hydrophobicity as compared to the wild-type LigIV. |
| Databáze: | OpenAIRE |
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