DNA-binding function of c-Myb R2R3 around thermal denaturation temperature
The minimum DNA-binding domain of the transcriptional factor c-Myb R2R3 remarkably fluctuates in the solution. In the present study, we evaluated the protein fluctuation of R2R3 C130I mutant, R2R3*, on its DNA-binding and folding thermodynamics. DNA-binding analysis using isothermal titration calor...
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| Format: | Article |
| Language: | English |
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The Biophysical Society of Japan
2021-04-01
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| Series: | Biophysics and Physicobiology |
| Subjects: | |
| Online Access: | https://doi.org/10.2142/biophysico.bppb-v18.009 |
| _version_ | 1818729275869626368 |
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| author | Maki Kawasaki Masayuki Oda |
| author_facet | Maki Kawasaki Masayuki Oda |
| author_sort | Maki Kawasaki |
| collection | DOAJ |
| description | The minimum DNA-binding domain of the transcriptional factor c-Myb R2R3 remarkably fluctuates in the solution. In the present study, we evaluated the protein fluctuation of R2R3 C130I mutant, R2R3*, on its DNA-binding and folding thermodynamics. DNA-binding analysis using isothermal titration calorimetry revealed that the heat capacity change determined from the correlation between temperature and binding enthalpy change is highly negative above 35°C, indicating that the fluctuation increases with increasing temperature and elevates the conformational change on DNA binding. The results were in accordance with those of differential scanning calorimetry, which revealed that the heat capacity corresponding to thermal denaturation gradually increased above 35°C, followed by the broad transition peak. In contrast, the transition peak of R2R3* in the DNA-bound state was sharper and larger than that in the DNA-unbound state. The fluctuating form could transform into lesser fluctuating form upon DNA binding, resulting in a larger enthalpy change for denaturation of R2R3* in the DNA-bound state. It should also be noted that R2R3* could specifically bind to DNA around thermal denaturation temperature. This would be due to proteins with numerous fluctuations. Moreover, we discuss specific and non-specific DNA binding accompanied by the conformational change between well-ordered and disordered forms of R2R3* observed around the denaturation temperature. |
| first_indexed | 2024-12-17T22:43:18Z |
| format | Article |
| id | doaj.art-a08167f3efa044e8bbafa49478843ed5 |
| institution | Directory Open Access Journal |
| issn | 2189-4779 |
| language | English |
| last_indexed | 2024-12-17T22:43:18Z |
| publishDate | 2021-04-01 |
| publisher | The Biophysical Society of Japan |
| record_format | Article |
| series | Biophysics and Physicobiology |
| spelling | doaj.art-a08167f3efa044e8bbafa49478843ed52022-12-21T21:29:52ZengThe Biophysical Society of JapanBiophysics and Physicobiology2189-47792021-04-011810.2142/biophysico.bppb-v18.009DNA-binding function of c-Myb R2R3 around thermal denaturation temperatureMaki Kawasaki0Masayuki Oda1Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Kyoto 606-8522, JapanGraduate School of Life and Environmental Sciences, Kyoto Prefectural University, Kyoto 606-8522, JapanThe minimum DNA-binding domain of the transcriptional factor c-Myb R2R3 remarkably fluctuates in the solution. In the present study, we evaluated the protein fluctuation of R2R3 C130I mutant, R2R3*, on its DNA-binding and folding thermodynamics. DNA-binding analysis using isothermal titration calorimetry revealed that the heat capacity change determined from the correlation between temperature and binding enthalpy change is highly negative above 35°C, indicating that the fluctuation increases with increasing temperature and elevates the conformational change on DNA binding. The results were in accordance with those of differential scanning calorimetry, which revealed that the heat capacity corresponding to thermal denaturation gradually increased above 35°C, followed by the broad transition peak. In contrast, the transition peak of R2R3* in the DNA-bound state was sharper and larger than that in the DNA-unbound state. The fluctuating form could transform into lesser fluctuating form upon DNA binding, resulting in a larger enthalpy change for denaturation of R2R3* in the DNA-bound state. It should also be noted that R2R3* could specifically bind to DNA around thermal denaturation temperature. This would be due to proteins with numerous fluctuations. Moreover, we discuss specific and non-specific DNA binding accompanied by the conformational change between well-ordered and disordered forms of R2R3* observed around the denaturation temperature.https://doi.org/10.2142/biophysico.bppb-v18.009binding thermodynamicsdna-binding proteinprotein fluctuationthermal stability |
| spellingShingle | Maki Kawasaki Masayuki Oda DNA-binding function of c-Myb R2R3 around thermal denaturation temperature Biophysics and Physicobiology binding thermodynamics dna-binding protein protein fluctuation thermal stability |
| title | DNA-binding function of c-Myb R2R3 around thermal denaturation temperature |
| title_full | DNA-binding function of c-Myb R2R3 around thermal denaturation temperature |
| title_fullStr | DNA-binding function of c-Myb R2R3 around thermal denaturation temperature |
| title_full_unstemmed | DNA-binding function of c-Myb R2R3 around thermal denaturation temperature |
| title_short | DNA-binding function of c-Myb R2R3 around thermal denaturation temperature |
| title_sort | dna binding function of c myb r2r3 around thermal denaturation temperature |
| topic | binding thermodynamics dna-binding protein protein fluctuation thermal stability |
| url | https://doi.org/10.2142/biophysico.bppb-v18.009 |
| work_keys_str_mv | AT makikawasaki dnabindingfunctionofcmybr2r3aroundthermaldenaturationtemperature AT masayukioda dnabindingfunctionofcmybr2r3aroundthermaldenaturationtemperature |