Enhanced basepair dynamics pre-disposes protein-assisted flips of key bases in DNA strand separation during transcription initiation
IR@IICB: CSIR-Indian Institute of Chemical Biology, Kolkata
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| Title |
Enhanced basepair dynamics pre-disposes protein-assisted flips of key bases in DNA strand separation during transcription initiation
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| Creator |
Roy, Neeladri Sekhar
Debnath, Subrata Chakraborty, Abhijit Chakraborty, Prasenjit Bera, Indrani Ghosh, Raka Ghoshal, Nanda Chakrabartia, Saikat Roy, Siddhartha |
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| Subject |
Structural Biology & Bioinformatics
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| Description |
Localized separation of strands of duplex DNA is a necessary step in many DNA-dependent processes,
including transcription and replication. Little is known about how these strand separations occur. The strand-separated E.coli RNA polymerase–promoter open-complex structure showed four bases of the non-template strand, the master base �11A, �7, �6 and +2, in a flipped state and inserted into protein pockets. To explore whether any property of these bases in the duplex state pre-disposes them to flipping, NMR studies were performed on a wild-type promoter in the duplex state. Measurement of relaxation times indicates that a limited number of base pairs, including the flipped ones, have faster opening rates than the rest. Molecular dynamics studies also show an inherently high dynamic character of the �11A:T base pair in the wild-type strand-paired state. In order to explore the role of the RNA polymerase in the flipping process, we have used 2-aminopurine as a fluorescent probe. Slower kinetics of the increase of 2-aminopurine fluorescence was observed with RNA polymerases containing several
mutant s70s. This may be interpreted as the protein playing an important role in enhancing the flipping
rate. These results suggest that flipping of �11A, and perhaps other flipped bases observed in the opencomplex,
is facilitated by its inherent proclivity to open-up with further assistance from the protein, thus leading to a strand-open state. Other DNA-based processes that require strand-separation may use similar pathways for strand separation. We conclude that not only basepair stability, but also dynamics may play an important role in the strand-separation.
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| Date |
2018
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Identifier |
http://www.eprints.iicb.res.in/2759/1/Phys.Chem.Chem.Phys.%2C2018.pdf
Roy, Neeladri Sekhar and Debnath, Subrata and Chakraborty, Abhijit and Chakraborty, Prasenjit and Bera, Indrani and Ghosh, Raka and Ghoshal, Nanda and Chakrabartia, Saikat and Roy, Siddhartha (2018) Enhanced basepair dynamics pre-disposes protein-assisted flips of key bases in DNA strand separation during transcription initiation. PHYSICAL CHEMISTRY CHEMICAL PHYSICS, 20. pp. 9449-9459. |
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| Relation |
http://dx.org.doi.10.1039/c8cp01119b
http://www.eprints.iicb.res.in/2759/ |
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