Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction
Clamp loaders are AAA+ ATPases that load sliding clamps onto DNA. We mapped the mutational sensitivity of the T4 bacteriophage sliding clamp and clamp loader by deep mutagenesis, and found that residues not involved in catalysis or binding display remarkable tolerance to mutation. An exception is a...
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2021-04-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/66181 |
| _version_ | 1811236844670550016 |
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| author | Subu Subramanian Kent Gorday Kendra Marcus Matthew R Orellana Peter Ren Xiao Ran Luo Michael E O'Donnell John Kuriyan |
| author_facet | Subu Subramanian Kent Gorday Kendra Marcus Matthew R Orellana Peter Ren Xiao Ran Luo Michael E O'Donnell John Kuriyan |
| author_sort | Subu Subramanian |
| collection | DOAJ |
| description | Clamp loaders are AAA+ ATPases that load sliding clamps onto DNA. We mapped the mutational sensitivity of the T4 bacteriophage sliding clamp and clamp loader by deep mutagenesis, and found that residues not involved in catalysis or binding display remarkable tolerance to mutation. An exception is a glutamine residue in the AAA+ module (Gln 118) that is not located at a catalytic or interfacial site. Gln 118 forms a hydrogen-bonded junction in a helical unit that we term the central coupler, because it connects the catalytic centers to DNA and the sliding clamp. A suppressor mutation indicates that hydrogen bonding in the junction is important, and molecular dynamics simulations reveal that it maintains rigidity in the central coupler. The glutamine-mediated junction is preserved in diverse AAA+ ATPases, suggesting that a connected network of hydrogen bonds that links ATP molecules is an essential aspect of allosteric communication in these proteins. |
| first_indexed | 2024-04-12T12:15:06Z |
| format | Article |
| id | doaj.art-e3ba2e767c774bfb9759eabd614c6afb |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T12:15:06Z |
| publishDate | 2021-04-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-e3ba2e767c774bfb9759eabd614c6afb2022-12-22T03:33:27ZengeLife Sciences Publications LtdeLife2050-084X2021-04-011010.7554/eLife.66181Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junctionSubu Subramanian0https://orcid.org/0000-0001-6095-7021Kent Gorday1Kendra Marcus2Matthew R Orellana3Peter Ren4Xiao Ran Luo5Michael E O'Donnell6John Kuriyan7https://orcid.org/0000-0002-4414-5477Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States; Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States; Biophysics Graduate Group, University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United StatesHoward Hughes Medical Institute, Rockefeller University, New York, United StatesDepartment of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States; Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States; Department of Chemistry, University of California, Berkeley, Berkeley, United States; Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, United StatesClamp loaders are AAA+ ATPases that load sliding clamps onto DNA. We mapped the mutational sensitivity of the T4 bacteriophage sliding clamp and clamp loader by deep mutagenesis, and found that residues not involved in catalysis or binding display remarkable tolerance to mutation. An exception is a glutamine residue in the AAA+ module (Gln 118) that is not located at a catalytic or interfacial site. Gln 118 forms a hydrogen-bonded junction in a helical unit that we term the central coupler, because it connects the catalytic centers to DNA and the sliding clamp. A suppressor mutation indicates that hydrogen bonding in the junction is important, and molecular dynamics simulations reveal that it maintains rigidity in the central coupler. The glutamine-mediated junction is preserved in diverse AAA+ ATPases, suggesting that a connected network of hydrogen bonds that links ATP molecules is an essential aspect of allosteric communication in these proteins.https://elifesciences.org/articles/66181bacteriophage T4AAA+ proteinsallosteric communicationreplisome |
| spellingShingle | Subu Subramanian Kent Gorday Kendra Marcus Matthew R Orellana Peter Ren Xiao Ran Luo Michael E O'Donnell John Kuriyan Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction eLife bacteriophage T4 AAA+ proteins allosteric communication replisome |
| title | Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction |
| title_full | Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction |
| title_fullStr | Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction |
| title_full_unstemmed | Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction |
| title_short | Allosteric communication in DNA polymerase clamp loaders relies on a critical hydrogen-bonded junction |
| title_sort | allosteric communication in dna polymerase clamp loaders relies on a critical hydrogen bonded junction |
| topic | bacteriophage T4 AAA+ proteins allosteric communication replisome |
| url | https://elifesciences.org/articles/66181 |
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